Does alveolar soft-part sarcoma exhibit skeletal muscle differentiation? An immunocytochemical and biochemical study of myogenic regulatory protein expression

There has been persistent controversy regarding the nature of cell differentiation in alveolar soft-part sarcoma (ASPS) since its first description in 1952. Some studies suggest that ASPS might represent an unusual variant of skeletal muscle tumor, Given the availability of new monoclonal antibodies to probe for skeletal muscle differentiation and the rapid advance in immunocytochemical techniques for deparaffinized, formalin-fixed tissue sections, we wished to test the proposed hypothesis that ASPS might represent a new type of rhabdomyosarcoma Twelve archival samples of ASPS were retrieved, and we investigated the expression of two myogenic regulatory proteins, MyoD1 and myogenin, as rvell as other muscle-associated proteins, using sensitive immunocytochemical techniques. Despite the presence of desmin immunostaining in six ASPSs, no tumors were positive for either muscle actin or myoglobin Most importantly, no specimen showed nuclear expression of MyoD1 or myogenin, In 11 tumors, however, there was considerable granular immunostaining in the tumor cell cytoplasm with the anti-MyoD1 monoclonal antibody 5.8A, a phenomenon observed in various nonmuscle normal and neoplastic tissues with this antibody, To analyze the exact nature of immunostaining of MyoD1 and desmin in ASPS, biochemical analyses using available fresh frozen tumor tissue were performed, Although a 53-kDa band was noted with antidesmin antibody on Western blot analysis, no specific protein band that corresponds to the 45-kDa MyoD1 was detected with antibody 5.8A. These results confirm the presence of desmin in ASPS but argue against authentic expression of MyoD1, They also suggest that the cytoplasmic immunostaining observed with anti-MyoD1 antibody 5.8A most likely represents a nonspecific cross-reaction with an unknown cytoplasmic antigen, Considering the master role that MyoD1 and myogenin play in skeletal muscle commitment and differentiation and the lack of expression of these two proteins in ASPS as determined immunocytochemically and biochemically, we think that the histogenesis of ASPS remains unknown.

Bootstrap confidence regions for canonical variates: Application to studies of evolutionary differentiation

Theory recently developed to construct confidence regions based on the parametric bootstrap is applied to add inferential information to graphical displays of sample centroids in canonical variate analysis. Problems of morphometric differentiation among subspecies and species are addressed using numerical resampling procedures.

Effects of Short-term Hyper- and Hypoprolactinaemia on Hormones of the Pituitary, Gonad and -Thyroid Axis and on Semen Quality in Male Beagles

Effects of a short-term hyper- and hypoprolactinaemia on serum concentrations of LH, testosterone and semen quality in six male Beagles were investigated. Blood samples were collected at 3-day intervals for 12 weeks. The time span was divided into five 3-week periods: pre-treatment, metoclopramide (MCP) treatment (0.2 mg/kg orally three times daily), cabergoline (CAB) treatment (5 mu g/kg orally once daily), post-treatment 1 and post-treatment 2. In the latter, only semen characteristics were evaluated. Semen parameters were analyzed once per week during the whole 15-week investigation time. At the end of each period, the effects of a single intravenous injection of thyrotropin-releasing hormone (TRH; 10 mu g/kg) on the secretion of prolactin (PRL), LH, testosterone, thyroid-stimulating hormone and thyroxine (T4) were investigated. Pre-treatment serum PRL concentration increased under MCP (p < 0.05), followed by a decrease under CAB administration (p < 0.05). Luteinizing hormone and testosterone concentrations were not affected. Except for straight-line sperm velocity, semen quality did not differ between collection periods. A single iv TRH injection induced a significant PRL increase at 20 min in all experimental periods except during CAB treatment. Luteinizing hormone and testosterone did not show clear TRH-related changes. Basic T4 levels were significantly reduced after CAB treatment ( p < 0.05). The results of the present study demonstrate that MCP-induced short-term hyperprolactinaemia in male beagles does not seriously affect the hypothalamo-pituitary axis and semen quality.

Chromosomal differentiation of Hyla nana and Hyla sanborni (Anura, Hylidae) with a description of NOR polymorphism in H-nana

Specimens of Hyla nana and Hyla sanborni from a syntopic population were studied cytogenetically. These species are morphologically very similar and are frequently misidentified, confused with each other. Both species had a diploid chromosome number, 2n = 30. However, the karyotypes of H. nana and H. sanborni differed considerably from each other in the number of submetacentric and telocentric chromosomes. The two species also differed in their primary NOR-bearing chromosomes (metacentric pair 13 in H. nana and telocentric pair 12 in H. sanborni). Additional nucleolus organizer regions (NORs) were detected by Ag-NOR staining and FISH in chromosome pairs 1, 5, 6, 12, and 14 in seven specimens of H. nana. Thus, a total of six patterns of NOR were identified. These differences in karyotype and in NOR location allowed the unambiguous identification of syntopic individuals of the two species. However, the chromosomal morphology of both species differed from that reported for populations from other geographic regions, suggesting that a systematic reevaluation of this group of Hyla may be necessary.

Differentiation of the honey bee (Apis mellifera L.) antennal lobes during metamorphosis: a comparative study among castes and sexes

In insects the antennal lobes (AL) constitute the brain deutocerebrum. In bees they consist of two neuropil regions, each associated with one antenna, delimited by a layer of glial cells and somata of neurons. The neuropil is organized in distinct globular structures of dense synaptic axons coming from the olfactory organs of the antennae, known as glomeruli. In Apis mellifera, as in other eusocial species of bees, queens, workers, and drones perform different functions in the colony and consequently the organs associated with these functions undergo a differential development. In this paper we analyzed the structure and size of the differentiating AL of queens, workers, and drones during metamorphosis using light microscopy. During metamorphosis the neuropil enlarge and differentiates into concentric structures known as glomeruli. The results showed size, structural and temporal differences in the glomeruli development among the classes of individuals of the colony. The neuropil differentiation starts early and is faster in drones and newly emerged worker is the colony individual class with greater neuropil area in AL. These results are discussed taking in account the functions of the individuals in the colony. (C) Koninklijke Brill NV, Leiden, 2011.

Conventional and Doppler ultrasound for the differentiation of benign and malignant canine mammary tumours

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Sex chromosome differentiation in Belostoma (Insecta: Heteroptera: Belostomatidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Allozyme differentiation among allopatric populations of Anopheles nuneztovari (Diptera: Culicidae)

Four enzymes in six geographic populations, four Brazilian and two Colombian, of Anopheles nuneztovari were studied. There were differences among the most frequent alleles in the EST5, ACON and MDH loci in the populations from Brazil and Colombia. The α-GPD(*)C allele was encountered very frequently in the Palo/Sit population in Colombia and the α-GPD(*)B allele was found to be fixed in most of the Brazilian populations. An additional α-GPD band was found only in Palo/Sit. The considerable genetic differentiation between populations in western Colombia vs. Brazil suggests that a certain degree of reproductive isolation exists between them.

Differentiation of floral color and odor in two fly pollinated species of Metrodorea (Rutaceae) from Brazil

We investigated if differences in morphological characters in two species of Metrodorea (Rutaceae) from Brazilian semideciduous forests correspond to some pollination divergence. M. nigra and M. stipularis are sympatric species, display a similar floral morphology, are protandrous, self-incompatible, their flower periods overlap, and both are pollinated by flies. M. nigra main pollinators are Pseudoptiloleps nigripoda (Muscidae) and Fannia sp. (Fanniidae); M. stipularis major pollinators are Phaenicia eximia (Calliphoridae), Palpada sp. and Ornidia obesa (Syrphidae). The distinct floral odor (disagreeable in M. nigra and sweet in M. stipularis) and color (brownish violet vs. pale yellow) determine the differences on type and number of floral visitors observed. Several species from semideciduous forests initially considered to be pollinated by diverse insects, present flies as main pollinators, stressing the importance of fly pollination in such habitats.

Subcutaneous and testicular metastasis from prostatic adenocarcinoma with neuroendocrine differentiation

The pathological finding of testicular metastasis in cases of disseminated prostatic adenocarcinoma is rare, but was more frequently reported in the past, when bilateral castration was performed more often. The existence of skin and subcutaneous metastasis adds a worse prognosis, because generally it is sign of advanced disease with an average survival time of less than one year. The synchronous occurrence of such metastasis has not been described previously, neither their association to neuroendocrine differentiation. The presence of such differentiation of prostatic adenocarcinoma represents a very unfavorable prognostic factor, as suggested in recent literature. Herein, we discuss the case of a 53 year old man, who presented with macroscopic hematuria and frequency associated to several painless subcutaneous nodules in left axilla and shoulder, as well as in the lower abdominal wall. The right testis was painful, endured and on rectal examination, the prostate was diffusely enlarged. Serum PSA was elevated, reaching 1760 ng/ml and prostatic biopsy disclosed a Gleason 10 prostatic adenocarcinoma with neuroendocrine differentiation. The same pathological pattern was detected in the right testis and in all subcutaneous nodules, documented by positive staining of chromogranin, a marker of neuroendocrine cells. He was submitted to a prostate tunnelization and maximal androgen blockade plus adjuvant chemotherapy, nevertheless, he died 5 months latter.

Mating influence in the ovary differentiation in adult queens of Apis mellifera L. (Hymenoptera, Apidae)

The present results show that in the ovarioles of a newly emerged (0 day) queen of A. mellifera only two regions may be distinguished: a proximal, short germarium and a very long distal, terminal filament. As the queen matures and gets ready for the nupcial flight, the germarium increases in lenght, advancing towered the distal end, as the terminal filament shortens. The ovarioles of queens ready to mate (6 to 8 days old) have, already one or two ovarian follicles, i.e. a very short proximal vitellarium, but a real vitellogenesis only starts after the fecundation. If the queen does not mate the ovarioles structure is disrupted (12-16 days old). In mated queen eggs the ovarioles present three differentiated regions, from the apice to the basis: a short terminal filament, a medium size germarium, and a very long basal vitellarium. As the eggs are laid, the emptied follicle collapses, degenerates and produces a corpus luteum.

Desenvolvimento morfolôgico dos ovários em embriões e fetos bovinos da raça Nelore

The morphologic events related to the prenatal development of the ovaries in 81 Nelore breed embryos and fetuses gathered in a local slaughterhouse, with age range from 26 to 240 days following fecundation were studied. The age of fetuses was estimated from measures taken in the cranium-caudal direction. The sex was identified from macroscopic observations and using Polymerase Chain Reaction (PCR) technique. For histology the gonads were fixed in Bourn's fluid for 24 hours and 5 μm thick section's were stained with hematoxylin-eosin. Formation of gonadal ridge and presence of germinal cells were found within it at 29 and 34 days, respectively. Oogonia and primordial follicles, unlike the growing follicles, exhibited significant differences in diameter in the various periods studied. Positive correlation (P<0.05) was found between the diameter of oogonia and their nucleus as well as between primordial and growing follicles with their oocytes and respective nuclei. The gonad was fully formed at 40 days. Primordial follicles, in the growing stage, and antral follicles first appeared, approximately at 95, 140, and 180 days, respectively. Despite the onset and duration of oogenesis being similar to that of taurine breeds, folliculogenesis initiates at an early stages in the Nelore breed.

A hydrogel scaffold that maintains viability and supports differentiation of dental pulp stem cells

Objectives: The clinical translation of stem cell-based Regenerative Endodontics demands further development of suitable injectable scaffolds. Puramatrix™ is a defined, self-assembling peptide hydrogel which instantaneously polymerizes under normal physiological conditions. Here, we assessed the compatibility of Puramatrix™ with dental pulp stem cell (DPSC) growth and differentiation. Methods: DPSC cells were grown in 0.05-0.25% Puramatrix™. Cell viability was measured colorimetrically using the WST-1 assay. Cell morphology was observed in 3D modeling using confocal microscopy. In addition, we used the human tooth slice model with Puramatrix™ to verify DPSC differentiation into odontoblast-like cells, as measured by expression of DSPP and DMP-1. Results: DPSC survived and proliferated in Puramatrix™ for at least three weeks in culture. Confocal microscopy revealed that cells seeded in Puramatrix™ presented morphological features of healthy cells, and some cells exhibited cytoplasmic elongations. Notably, after 21 days in tooth slices containing Puramatrix™, DPSC cells expressed DMP-1 and DSPP, putative markers of odontoblastic differentiation. Significance: Collectively, these data suggest that self-assembling peptide hydrogels might be useful injectable scaffolds for stem cell-based Regenerative Endodontics. © 2012 Academy of Dental Materials.