Immunohistochemical techniques and their applications in the histopathology of the respiratory system.

Subsequent to the first report in the 1940s on incubation of tissue sections with fluorescein-conjugated antibodies for localization of antigens, a great number of modifications were introduced to improve the validity of immunohistochemistry which has become a growingly popular tool. The use of immunoenzymatic techniques eliminates the need for expensive fluorescence microscopy equipment, the lack of permanency of preparations and the lack of electron density required in ultrastructural localization of antigens. Regardless of the technique, it is also important to choose a correct fixation which allows the proper preservation of antigens and morphology and the penetration of antibodies through the entire thickness of the preparation. A variety of immunohistochemical techniques have been applied to study several components of the lung, such as collagen, surface active material, lung specific antigens, and enzymes and the detection of tumor markers, immunoglobulins and infectious agents in the respiratory system which is reviewed. The large surface area and the multiplicity of cell types provided by the respiratory tract epithelium of humans for exposure to microbial as well as toxic substances in the environment make this organ system very vulnerable but a good early indicator of adverse health effects. Immunohistochemistry provides valuable information complementary to the immunochemical and biochemical characterization of this barrier

Microarray analysis of androgen-regulated gene expression in testis: the use of the androgen-binding protein (ABP)-transgenic mouse as a model

BACKGROUND: Spermatogenesis is an androgen-dependent process, yet the molecular mechanisms of androgens' actions in testis are poorly understood. Transgenic mice overexpressing rat androgen-binding protein (ABP) in their testes have reduced levels of intratesticular androgens and, as a result, show a progressive impairment of spermatogenesis. We used this model to characterize changes in global gene expression in testis in response to reduced bioavailability of androgens. METHODS: Total RNA was extracted from testes of 30-day old transgenic and wild-type control mice, converted to cRNA, labeled with biotin, and hybridized to oligonucleotide microarrays. Microarray results were confirmed by real-time reverse transcription polymerase chain reaction. RESULTS: Three-hundred-eighty-one genes (3.05% of all transcripts represented on the chips) were up-regulated and 198 genes (1.59%) were down-regulated by at least a factor of 2 in the androgen-deficient animals compared to controls. Genes encoding membrane proteins, intracellular signaling molecules, enzymes, proteins participating in the immune response, and those involved in cytoskeleton organization were significantly overrepresented in the up-regulated group. Among the down-regulated transcripts, those coding for extracellular proteins were overrepresented most dramatically, followed by those related to proteolysis, cell adhesion, immune response, and growth factor, cytokine, and ion channel activities. Transcripts with the greatest potential impact on cellular activities included several transcription factors, intracellular signal transducers, secreted signaling molecules and enzymes, and various cell surface molecules. Major nodes in the up-regulated network were IL-6, AGT, MYC, and A2M, those in the down-regulated network were IL-2, -4, and -10, MAPK8, SOCS1, and CREB1. CONCLUSION: Microarray analysis followed by gene ontology profiling and connectivity analysis identified several functional groups of genes and individual genes responding to sustained reduction of androgen levels in the mouse testis. These include genes whose products function as transcription factors, cell surface molecules including ion channels, extra- and intracellular signaling molecules, and secreted enzymes with the potential of regulating cell-to-cell attachment. The transcription factors CREB1 (down-regulated) and MYC (up-regulated) may mediate the most important initial phases of the testicular response to reduced levels of androgens. These results suggest specific avenues for further research that will lead to a better understanding of how androgens regulate spermatogenesis

Dynamics of testicular germ cell apoptosis in normal mice and transgenic mice overexpressing rat androgen-binding protein

The number and type of testicular germ cells undergoing apoptosis in different age groups of mice (from 7 to 360 days of age) was determined and compared in age-matched wild type (WT) control and in a transgenic (TG) mice homozygous to rat androgen binding protein (ABP) using flow cytometry. Flow cytometric quantification revealed that the total number of germ cells undergoing apoptosis did not differ significantly in WT and TG mice up to Day 14. From Day 21 to Day 60, the number of germ cells undergoing apoptosis was consistently higher in TG than in WT mice. Starting from Day 90, the number of germ cells undergoing apoptosis in TG mice was lower than controls until Day 360. In 21–60 days old TG mice, spermatogonia, S-Phase cells, and primary spermatocytes are the cell types undergoing apoptosis at significantly greater numbers than those in WT mice. However, starting from day 60, the total number of spermatids undergoing apoptosis was significantly lower in TG mice than in age-matched WT controls. TdT-mediated dUTP-biotin nick end labeling (TUNEL) in testicular sections from TG mice of 21 and 30 days of age confirmed the presence of increased numbers of apoptotic germ cells compared to their age matched controls. These data indicate that the continuous presence of greater than physiological concentrations of ABP in the mouse testis has a biphasic effect on the frequency of apoptosis in germ cells. The initial pre-pubertal increase in testicular germ cell apoptosis may result from direct or indirect actions of ABP and is likely to determine the subsequent life-death balance of germ cell populations in TG mice, whereas the subsequent reduction may result from maturation depletion. A wave of apoptosis during the pre-pubertal period is required for normal spermatogenesis to develop, and our data indicate that this apoptotic wave may be regulated by ABP and/or androgens

EVALUASI PENYELENGGARAAN PROGRAM PEMBINAAN CABANG OLAHRAGA TAEKWONDO DI SEKOLAH KHUSUS OLAHRAGA (SKO) RAGUNAN

Penelitian ini bertujuan untuk mendapatkan informasi tentang penyelenggaraan Program Pembinaan Cabang Olahraga Taekwondo di Sekolah Khusus Olahraga (SKO) Ragunan Jakarta. Penelitian evaluasi program ini dilaksanakan dalam rangka menguji tingkat ketercapaian tujuan-tujuan yang telah ditetapkan pada penyelenggaraan program Pembinaan cabang olahraga Taekwondo. Penelitian ini dilaksanakan di Jakarta pada Asisten Deputi Pengelolaan Sentra Keolahragaan Bidang Sekolah Khusus Olahraga Deputi III Pembudayaan Olahraga Kementerian Pemuda dan Olahraga Republik Indonesia, beralamat di Jalan Gerbang Pemuda No. 3 Senayan Jakarta Pusat. Penelitian ini dilaksanakan pada bulan Januari sampai dengan Maret 2020. Penelitian evaluasi ini metode penelitian kualitatif dengan pendekatan model evaluasi Context, Input, Process, Product (CIPP). Pengumpulan data dilakukan dengan cara observasi, wawancara dan studi dokumentasi. Kesimpulan dari hasil penelitian yaitu: Tahapan context, penyelenggaraan program pembinaan cabang olahraga Taekwondo di Sekolah Khusus Olahraga (SKO) Ragunan Jakarta dikategorikan sangat baik dan terdapat kesesuaian dalam pelaksanaan program dengan visi dan misi yang hendak dicapai. Tahapan Input, berdasarkan hasil evaluasi dari lima komponen yang meliputi sistem perekrutan atlet, sistem perekrutan pelatih, sarana dan prasarana, pembiayaan dan prosedur program latihan diperoleh hasil cukup baik dengan pedoman penyelenggaraan program pembinaan cabang olahraga taekwondo. Tahapan proses meliputi proses pelaksanaan dan peran tim monitoring dan evaluasi program latihan dikategorikan baik. Tahapan produk yang di evaluasi adalah yakni prestasi atlet. evaluasi ini sudah sesuai harapan dari penyelenggaraan program pembinaan cabang olahraga Taekwondo di Sekolah Khusus Olahraga (SKO) Ragunan Jakarta. Kata Kunci: Evaluasi Penyelenggaraan Program Pembinaan Cabang Olahraga Taekwondo di Sekolah Khusus Olahraga (SKO) Ragunan. *************** The purpose of this purpose is to get an information about Taekwondo menegement founding programme in Jakarta sepecial sport school. Evalution research of this programme is done in the way to test an achievement level of purpose that had been determined in the Taekwondo founding programme. This research was done in Jakarta to the Central superintendence Sport Deputy Assitant for special sport school sector Deputy III on sports cultivation in Young and Sports Ministry of Republic Indonesian which addressed at Jalan Gerbang Pemuda Number 3 senayan, Central Jakarta. It was done on January up to March 2020. This Evaluation research uses a qualitative method that are using model evaluation approach context, Input, Process, Product (CIPP). The Collection Data was done by observation, interview, and documentation study. The result of research conclusion are contain of some stages like first, context stage, it is about Taekwondo management founding programme in Jakarta Ragunan special sport school which is categorized as a very good school and suitable with the vision and mission of the programme implementation that will be reached. Second is Input stag; based on five components that are contain of athlete recruitment system, trainer recruitment system, facilities and infrastructure, Financing and procedure training programme had gotten with the good enough result based on Taekwondo management founding programme. Fourt, process stage that include implementation process, monitoring and evaluation exercise programme teams are good categoried. Last product stage that evaluated about athlete achievement. Those evaluations are have suitable with an expectation from Taekwondo management programme in Jakarta Ragunan special sport school. Key Word : “Evaluation of Taekwondo Management Programme in Jakarta Ragunan Special Sport School”

Hubungan Antara Citra Toko (Store Image) Toko Pernak-Pernik dan Aksesoris dengan Perilaku Membeli Secara Impulsif (Impulsive Buying) pada Konsumen Remaja Putri

Saat ini kebutuhan dan keinginan seseorang akan memiliki suatu barang akan membuatnya menjadi konsumen yang selalu ingin memenuhi kebutuhannya. Fenomena yang terjadi adalah adanya kebutuhan dan keinginan yang dimiliki oleh wanita terhadap fashion dan tren-tren lain yang mendukung seseorang konsumen remaja putri untuk membeli suatu produk tanpa mempertimbangkan faktor perlu atau tidaknya produk tersebut. Produk fashion dan aksesoris sebagai penunjang penampilan fisik seseorang merupakan produk yang paling banyak diminati oleh para konsumen remaja putri. Toko pernak-pernik yang ada seperti Stroberi, Naughty, How R U, Bunga, dsb menjadi beberapa pilihan yang sedang menjamur saat ini dan digemari oleh kalangan para remaja putri sebagai tempat untuk membeli berbagai pernak-pernik dan aksesoris. Saat remaja putri terdorong untuk membeli dan selanjutnya membeli tanpa adanya perencanaan maka muncullah perilaku impulsive buying. Banyak faktor yang mempengaruhi munculnya perilaku ini namun penelitian ini akan melihat dari faktor toko pernak-pernik itu sendiri. Tujuan dari penelitian ini ingin mengetahui apakah ada hubungan antara citra toko (store image) pernak-pernik dan aksesoris tersebut dengan munculnya perilaku membeli secara impulsif terhadap produk toko pada diri remaja putri sebagai target konsumen dari toko tersebut. Subjek penelitian ini berjumlah 83 orang yang terdiri dari mahasiswi dan siswi SMU sebagai konsumen toko pernak­ pernik dan aksesoris. Teknik yang digunakan adalah acccidental sampling. Pengambilan data dilakukan dengan menyebar angket terbuka dan tertutup dengan skala Likert. Data yang ada selanjutnya diolah dengan menggunakan korelasi Product-Moment. Hasilnya adalah ada hubungan positif antara citra toko (store image) toko pernak-pernik dengan perilaku impulsive buying dengan nilai koefisien sebesar 0.307 dengan nilai sig. 0.005, yang berarti semakin tingi citra toko pernak-pernik yang dimiliki oleh remaja putri maka semakin tinggi perilaku impulsive buying. Sebanyak 51.8% subjek penelitian memiliki citra toko pernak-pernik dan aksesoris yang tinggi dan sebanyak 42.2% subjek memiliki perilaku impulsive buying yang tinggi. Penelitian ini juga menemukan beberapa faktor lain yang dapat mempengaruhi munculnya perilaku impulsive buying, yaitu frekuensi subjek pergi ke toko pernak-pernik, ada atau tidaknya perencanaan sebelum berbelanja serta dengan siapa subjek berbelanja

Identification, cloning and functional characterization of novel sperm associated antigen 11 (SPAG11) isoforms in the rat

BACKGROUND: Sperm binding proteins and their C-terminal peptides of the Sperm Associated Antigen 11 (SPAG11) family were found to play an important role in epididymal innate immunity in addition to their role in sperm maturation. However, the expression of Spag11 transcripts in rodents is not well documented. METHODS: Computational analysis was employed to identify novel Spag11 isoforms in the rat. RT-PCR analyses were carried out on RNAs isolated from the male reproductive tract tissues of rat using gene specific primers for Spag11c and Spag11t. The identities of PCR products were confirmed by sequencing. Tissue distribution, developmental expression and androgen regulation of Spag11t and Spag11c were studied using RT-PCR. The antimicrobial activities of recombinant Spag11t and Spag11c were tested against E coli in a colony forming unit assay. RESULTS: In this study, we identified two novel Spag11 transcripts, namely, Spag11t and Spag11c derived from the long arm of chromosome 16 in the rat (Rattus norvegicus), using both in silico and molecular biology approaches. Spag11c is expressed in all three regions of the epididymis, in testis and in ovary but is absent from the seminal vesicle. Spag11t expression is confined to the caput and it is not expressed in the testis, seminal vesicle or ovary. Age dependent expression of Spag11t and Spag11c was observed in the epididymides of rats (10–60 day old). Their expression was found to be most abundant in the adult rat (60 day) suggesting roles in mature reproductive function. Further, both Spag11t and Spag11c expression was down regulated in castrated rat epididymides and the expression was maintained in the testosterone replaced castrated rats. SPAG11C is a potent antibacterial agent. SPAG11T also displayed bactericidal capacity although weaker than SPAG11C and SPAG11E. CONCLUSION: The abundant expression of Spag11t and Spag11c in the male reproductive tract suggests an important role in male reproductive tract immunity. Their expression is developmentally regulated and androgen dependent. Characterization of novel SPAG11 isoforms will contribute to our understanding of the role of epididymal proteins in sperm maturation and innate immunity

Characterization and functions of beta defensins in the epididymis

The epididymal beta-defensins have evolved by repeated gene duplication and divergence to encode a family of proteins that provide direct protection against pathogens and also support the male reproductive tract in its primary function. Male tract defensins also facilitate recovery from pathogen attack. the beta-defensins possess ancient conserved sequence and structural features widespread in multi-cellular organisms, suggesting fundamental roles in species survival. Primate SPAG11, the functional fusion of two ancestrally independent beta-defensin genes, produces a large family of alternatively spliced transcripts that are expressed according to tissue-specific and species-specific constraints. the complexity of SPAG11 varies in different branches of mammalian evolution. Interactions of human SPAG11D with host proteins indicate involvement in multiple signaling pathways.Univ N Carolina, Reprod Biol Lab, Chapel Hill, NC 27599 USAPondicherry Cent Univ, Dept Biochem & Mol Biol, Pondicherry 605014, IndiaUniversidade Federal de São Paulo, Dept Pharmacol, Sect Expt Endocrinol, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Pharmacol, Sect Expt Endocrinol, BR-04044020 São Paulo, BrazilWeb of Scienc

Preferential immunohistochemical localization of vasoactive intestinal polypeptide (VIP) in the sacral spinal cord of the cat: light and electron microscopic observations

In the present study we have employed immunoperoxidase techniques to investigate the distribution of vasoactive intestinal polypeptide (VIP)- like immunoreactivity in the spinal cord and sensory ganglia of the cat. The spinal distribution of VIP-containing neuronal processes was also compared with that of substance P (SP), somatostatin (SOM), and cholecystokinin-8 (CCK) at lumbar, sacral, and coccygeal levels. At sacral levels, VIP was found to be contained in small and medium-sized primary sensory neurons and in dorsal rootlets. Deafferentation, by either ganglionectomy or dorsal rhizotomy, resulted in a nearly complete loss of VIP immunoreactivity in the spinal cord. The spinal distribution of VIP fibers and terminals was most dense and extensive in sacral segments. Forming a thin shell around the dorsal horn, collaterals, apparently originating from Lissauer's tract, projected either medially or laterally through lamina I. Laterally, many VIP axons terminated in lateral laminae V to VII. Others projected further through the neck of the dorsal horn to medial lamina V and the gray matter near the central canal. Medially, VIP axons descended through lamina I to expand into terminal fields in the posterior commissure and medial lamina V. At the ultrastructural level, VIP-like immunoreactivity was found in dense core vesicles within axonal enlargements containing both large dense core and smaller clear round vesicles. Synaptic connections were infrequently observed but, when encountered, were of the simple axodendritic type. The spinal distribution of VIP-containing fibers was remarkably similar to that reported for pelvic nerve visceral afferents, both in termination patterns within the spinal gray matter and in localization to the sacral cord. The density of SP-, SOM-, and CCK-containing fibers and terminals was constant at all levels examined (L4 to Co4). In marked contrast, the distribution of VIP fibers, much like that of pelvic nerve afferents, was mostly confined to sacral segments. Thus, although SP, SOM, and CCK may be contained within a population of sacral visceral afferents, they must be common to afferent systems in other segments as well. VIP, however, appears to be preferentially contained within pelvic visceral afferent fibers confined mostly to sacral segments

Localization of androgen-binding protein in proliferating Sertoli cells in culture.

The peroxidase and immunofluorescent localization patterns of androgen-binding protein (ABP), a biological marker of Sertoli cell function, have been examined in cultured Sertoli cells isolated from 20- to 22-day-old rats. ABP immunoreactivity in the form of cytoplasmic granules of variable diameter was observed in Sertoli cells with characteristic lipid droplets and a colony-forming, epithelial-like growth pattern. Incubation of cultures with [3H]thymidine demonstrated that Sertoli cells continue to produce ABP while retaining their capability for synthesizing DNA and undergoing mitosis. A variable number of cultured Sertoli cells became morphologically transformed after exposure to follitropin (follicle-stimulating hormone) and pharmacological agents acting on cyclic nucleotide metabolism. The induced change in Sertoli cell shape coincided with a disappearance of ABP-containing granules from the cytoplasm. These observations demonstrate that localization of ABP by immunological techniques is a valuable tool for the characterization of structural and functional properties of Sertoli cell in culture