Histochemistry

Based on a "field-effect" theory in colon carcinogenesis, and the expression of the disaccharide tumor marker D galactose-beta-[1-->3]-N-acetyl-D-galactosamine (Gal-GalNAc) in the rectal mucus of patients with cancer and precancer of the colon, Shamsuddin developed a simple, accurate, inexpensive, easy to perform and rapid (< or = 15 min) screening test for colonic cancer and precancerous lesions. In this study we examined 137 rectal mucus samples of randomly selected patients with colorectal malignancy or other colorectal diseases to confirm the sensitivity and specificity of this test in Croatia. Additionally, to test the validity of the "field-effect" theory, that the mucosa away from the obvious cancer will show abnormalities as a result of the generalized effect of the carcinogen throughout the entire field of the target tissue, we also monitored a subset of 53 patients post-operatively. Individuals free of colonic or any other malignancies served as c...

The cell bodies of pseudounipolar neurons in the trigeminal ganglia carry sensation from the face to the brainstem. These neurons vary in its sizes that carry specific sensations. In this study, trigeminal ganglion neurons are classified based on its sizes in to three types namely large, medium and small sized with the help of histomorphometry. Both side trigeminal ganglia showed more frequency (48%) of medium sized neurons which carry all types of sensation like pain, touch etc. The right side trigeminal ganglia showed more number of large sized neurons than left side ganglia.

Glutamine synthetase (GS) has long been known to be expressed exclusively in pericentral hepatocytes most proximal to the central veins of liver lobuli. This enzyme as well as its peculiar distribution complementary to the periportal compartment for ureogenesis plays an important role in nitrogen metabolism, particularly in homeostasis of blood levels of ammonium ions and glutamine. Despite this fact and intensive studies in vivo and in vitro, many aspects of the regulation of its activity on the protein and on the genetic level remained enigmatic. Recent experimental advances using transgenic mice and new analytic tools have revealed the fundamental role of morphogens such as wingless-type MMTV integration site family member signals (Wnt), beta-catenin, and adenomatous polyposis coli in the regulation of this particular enzyme. In addition, novel information concerning the structure of transcription factor binding sites within regulatory regions of the GS gene and their interactions with signalling pathways could be collected. In this review we focus on all aspects of the regulation of GS in the liver and demonstrate how the new findings have changed our view of the determinants of liver zonation. What appeared as a simple response of hepatocytes to blood-derived factors and local cellular interactions must now be perceived as a fundamental mechanism of adult tissue patterning by morphogens that were considered mainly as regulators of developmental processes. Though GS may be the most obvious indicator of morphogen action among many other targets, elucidation of the complex regulation of the expression of the GS gene could pave the road for a better understanding of the mechanisms involved in patterning of liver parenchyma. Based on current knowledge we propose a new concept of how morphogens, hormones and other factors may act in concert, in order to restrict gene expression to small subpopulations of one differentiated cell type, the hepatocyte, in different anatomical locations. Although many details of this regulatory network are still missing, and an era of exciting new discoveries is still about to come, it can already be envisioned that similar mechanisms may well be active in other organs contributing to the fine-tuning of organ-specific functions.

Breast cancer is a major health problem in underdeveloped countries including Sudan. Immunohistochemical typing is important for diagnosing and understanding the biology of such heterogeneous disease. To the best of our knowledge, no study has investigated the association between estrogen receptors (ER) expression and carbohydrates in breast tissue. Mammographic high density, a major risk factor, is partially attributed to carbohydrates. This study aimed to determine the type of carbohydrates in malignant and benign breast lesions and detect if there is any association between ER expression and carbohydrates using histochemical and immunohistochemical methods. A total of 60 diagnosed samples of different breast lesions were reexamined for ER status by immunohistochemistry (IHC) and for carbohydrates by histochemical staining methods i.e. periodic acid Schiff (PAS) and alcian blue. Strong relationship was discovered between ER positivity and presence of acid mucin, but not with sulfated mucin. Furthermore, strong relationship was found between ER positivity and PAS positive materials, but not with glycogen. Since there was a significant correlation between ER and carbohydrates, these positive PAS materials might be PAS positive acid mucin. These findings are useful in understanding the behavior of breast lesions. Also, these findings suggested an underlying molecular relationships and that ER expression might be predicted using histochemical demonstration of carbohydrates in female breast tissue particularly when immunohistochemistry is out of reach.

The somitomeres are spherical clusters of the paraxial mesenchymal cells. They were demonstrated only by the scanning electron microscope. No light microscopic criteria were described for the developing somitomeres. In this study the lectin binding pattern of wheat germ agglutinin and succinylated wheat germ agglutinin was used to verify the histochemical specification of the segment identity in view of the previously described scheme of somitogenesis in chick embryos. The results  showed that recruitment of the paraxial mesodermal cells is acompanied by localization of N-acetylglucosamine; organization of these cells occured in association with intracellular accumulation of sialic acid, while the segmentation of the somites was correlated with diffusion of sialic acid in the paraxial mesodermal tissue.

Intestinal metaplasia (IM) is part of a stepwise sequence of alterations of the gastric mucosa, leading ultimately to gastric cancer, and is strongly associated with chronic Helicobacter pylori infection. The molecular mechanisms underlying the onset of IM remain elusive. The aim of this study was to assess the putative involvement of two intestine-specific transcription factors, CDX1 and CDX2, in the pathogenesis of gastric IM and gastric carcinoma. Eighteen foci of IM and 46 cases of gastric carcinoma were evaluated by immunohistochemistry for CDX1 and CDX2 expression. CDX1 was expressed in all foci of IM and in 41% of gastric carcinomas; CDX2 was expressed in 17/18 foci of IM and in 54% of gastric carcinomas. In gastric carcinomas, a strong association was observed between the expression of CDX1 and CDX2, as well as between the intestinal mucin MUC2 and CDX1 and CDX2. No association was observed between the expression of CDX1 and CDX2 and the histological type of gastric carcinoma. In conclusion, these results show that aberrant expression of CDX1 and CDX2 is consistently observed in IM and in a subset of gastric carcinomas. The association of CDX1 and CDX2 with expression of the intestinal mucin MUC2, both in IM and in gastric carcinoma, indirectly implies that CDX1 and CDX2 may be involved in intestinal differentiation along the gastric carcinogenesis pathway. Copyright © 2002 John Wiley & Sons, Ltd.

A comparative histomorphological study was conducted on thymus of broiler chicken,
duck and quail. The thymus of all three species of post-hatch (adult) birds showed a thin connective
tissue capsule. However, fine septa originating from the capsule divided the organ into incomplete but
distinct lobules in chicken, whereas lobules were ill defined in both quail and duck. Each lobule, in
turn, was composed of a peripheral cortex, densely studded with lymphocytes and a central medulla
that was enriched with epithelial reticular cells. So, the cortex appeared more basophilic than that of
medulla. But the concentration of lymphocyte, an immune competent cell was highest in chicken
followed by duck and quail. Hassall’s corpuscles like structures were found inside the medulla as pale
stained diffuse body. General histo-architecture was almost similar in all the birds.

ABSTRAK Metode pewarnaan jaringan untuk mendeteksi adanya senyawa kimia di dalam jaringan salah satunya yaitu uji histokimia. Uji ini memerlukan reagen pewarna yang sesuai dengan senyawa yang ingin dideteksi. Praktikum ini bertujuan untuk mendeteksi keberadaan senyawa fitokimia pada bagian tertentu dari beberapa taksa tumbuhan dikotil dan monokotil. Sampel yang digunakan adalah umbi singkong, ubi jalar ungu, ubi kentang, air jagung, dan air kentang untuk uji amilum. Irisan jagung untuk uji selulosa, tempurung kelapa untuk uji lignin, daun kenanga dan daun jeruk untuk uji lipid, daun jambu untuk uji tanin, serta ubi jalar ungu dan jagung untuk uji gula reduksi. Metode yang dilakukan meliputi pembuatan preparat, pewarnaan, dan pengamatan. Pembuatan preparat dilakukan dengan membuat irisan sampel setipis mungkin ±5 µm menggunakan clamp on hand microtome, diletakkan di atas slide glass, ditetesi reagen pewarna spesifik, dan ditutup cover glas. Uji amilum menggunakan reagen lugol, gula pereduksi menggunakan reagen fehling diikuti proses pemanasan, selulosa menggunakan lugol dan H 2 SO 4 , lignin menggunakan reagen phloroglucin, lipid menggunakan reagen sudan III, serta tanin menggunakan FeCl 3 3% dan K 2 Cr 2 O 7 1%.

Natural products are studied to combat reproductive alterations of Lead. The current work aim to disclose the efficacy of Chlorella vulgaris and Zingiber officinale to alleviate lead acetate induced toxicity. Sixty adult male Wistar rats were distributed into four groups. Group 1 was considered control, group 2 received 200 mg/l PbAc water, group 3 received 50 mg/kg/rat of C. vulgaris extract and 200 mg/l PbAc water, and group 4 received 100 mg/kg/rat of Z. officinale and 200 mg/l PbAc water for 90 days. Testis samples were subjected to ultrastructural examination. It is observed that PbAc caused degenerative alterations in the spermatogenic series in many tubules, with a loss of germ cells and vacuoles inside the cytoplasm and between the germ cells. Mitochondria exhibited ballooning, with lost cristae and widening of the interstitial tissue, destructed nuclear envelopes of primary spermatocytes, and distortion of the axonemes of the mid-pieces of the sperms. With the treatment with C. vulgaris or Z. officinale, there were noticeable improvements in those modifications. It has concluded that both C. vulgaris and Z. officinale represent convincing medicinal components that may be used to ameliorate testicular toxicity in those exposed to lead in daily life with superior potentials revealed by C. vulgaris due to its chelating action.

The antiapoptotic, antioxidant, proliferative, and angiogenic effects of metallothionein (MT)-I+II has resulted in increased focus on their role in oncogenesis, tumor progression, therapy response, and patient prognosis. Studies have reported increased expression of MT-I+II mRNA and protein in various human cancers; such as breast, kidney, lung, nasopharynx, ovary, prostate, salivary gland, testes, urinary bladder, cervical, endometrial, skin carcinoma, melanoma, acute lymphoblastic leukemia (ALL), and pancreatic cancers, where MT-I+II expression is sometimes correlated to higher tumor grade/stage, chemotherapy/radiation resistance, and poor prognosis. However, MT-I+II are downregulated in other types of tumors (e.g. hepatocellular, gastric, colorectal, central nervous system (CNS), and thyroid cancers) where MT-I+II is either inversely correlated or unrelated to mortality. Large discrepancies exist between different tumor types, and no distinct and reliable association exists between MT-I+II expression in tumor tissues and prognosis and therapy resistance. Furthermore, a parallel has been drawn between MT-I+II expression as a potential marker for prognosis, and MT-I+II's role as oncogenic factors, without any direct evidence supporting such a parallel. This review aims at discussing the role of MT-I+II both as a prognostic marker for survival and therapy response, as well as for the hypothesized role of MT-I+II as causal oncogenes.

The main autocrine/paracrine role of the active metabolite of vitamin D(3), 1alpha,25-dihydroxyvitamin D(3) (1,25-D(3)), is inhibition of cell growth and induction of cell differentiation and/or apoptosis. Synthesis and degradation of the secosteroid occurs not only in the kidney but also in normal tissue or malignant extrarenal tissues such as the colon. Because 25-hydroxyvitamin D(3) 24-hydroxylase (CYP24A1) is considered to be the main enzyme determining the biological half-life of 1,25-D(3), we have examined expression of the CYP24A1 mRNA (by real-time RT-PCR) and protein (by immunohistochemistry) in normal human colon mucosa, colorectal adenomas, and adenocarcinomas in 111 patients. Although 76% of the normal and benign colonic tissue was either completely devoid of or expressed very low levels of CYP24A1, in the majority of the adenocarcinomas (69%), the enzyme was present at high concentrations. A parallel increased expression of the proliferation marker Ki-67 in the same sam...

The detailed characterization of multilayered Asian lacquer coatings is an important task for conservators in order to develop informed conservation and restoration approaches for complex Asian objects equipped with such decorative coatings. Of particular interest is often the identification of organic binders and their distribution within the layer structure. These days, the binding media contained in the individual lacquer layers are usually identified with sophisticated analytical equipment that is capable of great detail and precision, such as Fourier-transform infrared spectroscopy or pyrolysis-gas chromatography/mass spectrometry (PyGC/MS). However, the downside of using these techniques is that they are costly and not always accessible for many conservators, particularly at times when resources are tight. Alternative, low-tech methods that can be executed by the conservator himself are indeed available, but they are often deemed rather imprecise and potentially unreliable. Th...

Glandular trichomes of Labiatae are among the most investigated secretory structures. Most species studied belong to subfamily Nepetoidae, including plants with aromatic properties, while so far a few species of subfamily Lamioideae were examined. In this work, we studied the micromorphology, ultrastructure, type and release of secretion of the glandular trichomes present on leaves and flowers of several species belonging to subfamily Lamioideae, (Stachys alopecuros (L.) Bentham subsp. alopecuros, S. officinalis (L.) Trevisan subsp. officinalis, S. germanica L. subsp. germanica, S. germanica L. subsp. salviifolia (Ten.) Gams, S. sylvatica L., S. heraclea All., S. plumosa Griseb., S. annua L., Prasium majus L., Sideritis romana L.) and one to the sister group Scutellarioideae (Scutellaria galericulata L.). Besides the well-known peltate and small capitate trichomes, widely described in the literature, other types of glandular trichomes were encountered; stalked peltate hairs and large capitate hairs. In particular, a new type of capitate trichome, exclusive of calices and corollas, which presents a mode and release of secretion never described before, is reported.

We present a review of galanin in the brain from a historical perspective of the development of "chemoarchitectonics" and "brain cartography" accomplished in the Histopharmacology Section at the National Institutes of Health. It was the mapping of potential brain neuroregulators that served as a springboard of ideas from which behavioral studies emanate. The integration of the known localization of neurotransmitter/neuromodulatory nerves ("chemoarchitectonic maps") and receptor binding sites with biochemical data derived from brain micropunches coupled with behavioral analysis at the level of discrete brain allows one to define the anatomical circuits which support behavioral changes and which ultimately will improve our understanding of mental disorders.

Fishes have a great variety of structural and physiological adaptations that allow them to capture, digest and absorb food. In Hippocampus ingens it has been reported that they do not have teeth or stomach. However, it has the same intestinal structure as in other teleosts. In the present work the histological, histochemical characteristics and the presence of digestive enzymatic activity of the digestive tract of H. ingens are described with the purpose of providing information that can be used in the development of the culture of this species. We obtained 15 juveniles from a farm in Mazatlan, Sinaloa, Mexico. Some of them were fixed in the Davidson´s solution, sectioned at 4 μm using a rotatory microtome and stained with H-E, Alcian Blue-PAS, Toluidine Blue, Masson's Trichrome, Black Sudan and Ninhydrin-Schiff. To the rest, the digestive tract and the accessories were extracted to determine the presence of trypsin activity, chymotrypsin, lipase, amylase, aminopeptidase, acid phosphatase and alkaline phosphatase. The presence of an anterior and posterior esophagus was confirmed. Only the anterior region presents numerous acid-type mucosal cells, while the posterior region has simple cylindrical epithelium. Based on the type of epithelium, presence and height of villi as well as type and coverage of mucins present, the intestine differentiates into three regions. The anterior region presents acidic and neutral mucins, whereas in the middle and posterior regions it presents acidic mucins in greater production. The presence of vacuoles containing triglycerides within the enterocytes of the anterior intestine was recorded, as well as the proteins in the hepatocytes. The activity of aminopeptidase, chymotrypsin and amylase showed very low values with respect to lipase and alkaline phosphatase, which were the highest. Trypsin and acid phosphatase showed intermediate values. These results confirm that H. ingens presents an alkaline intestinal digestion and that it has high lipid requirements

Psidium guineense, commonly known as “araçá”, is a traditional medicinal plant used to treat diarrhea. The present study examined the morphoanatomy, histochemistry and phytochemistry of P. guineense leaves to evaluate the presence and localization of pharmacologically active compounds. The anatomical study employed traditional light microscope techniques. Histochemical and phytochemical tests and thin-layer chromatography (TLC) were used to detect flavonoids, tannins and essential oils. The leaves of P. guineense show characteristics common to other species of Myrtaceae, such a marginal vein, a brochidodromous enervation pattern, simple tector trichomes, paracitic stomata, and secretory cavities. Histochemical tests indicated the occurrence of flavonoids and tannins in the mesophyll and vascular system, and flavonoids and essential oils in secretory cavities. Phytochemical tests confirmed the presence of these compounds. The TLC demonstrated the diversity of the chemical substances revealed by many Rfs for the different groups of compounds evaluated. The knowledge of the morpho-anatomical characteristics of P. guineense can aid botanical diagnoses of this species, which has secondary compounds of therapeutic application.

The cellular processes involved in metal metabolism in molluscs are reviewed, with emphasis on the contribution of microscopy (AMG, ARG, EPMA, and SIMS) to both basic research of metal cell biology and applied environmental research. In molluscs, metal uptake may occur by facilitated diffusion, active transport, or endocytosis, and can be enhanced by MT synthesis or formation of mineralized granules. In aquatic molluscs, gills constitute a key interface for dissolved metal uptake, where metals are bound to MT, incorporated into lysosomes, and released basally towards the blood plasma and circulating hemocytes. However, particulate metal uptake is mainly achieved via the digestive tract by endocytosis; further metals are transferred first to lysosomes and then to residual bodies, especially in the digestive cells of the digestive gland. Additionally, metals can be accumulated selectively in specific cell types. As ligands pools differ from cell to cell, different metals may be retained in different cell types. Class “a” metals are localized in cells with granules composed of carbonate, oxalate, phosphate, and sulfate (oxygen donors), whereas “b” metals are associated with those cell types rich in sulfur and nitrogen ligands (sulfur donors). In molluscs, oxygen donors occur in connective tissue calcium cells and basophilic cells, whereas sulfur donors are present in digestive cells, podocytes, nephrocytes, and rhogocytes. Hemocytes, which constitute the most relevant system for metal transport between tissues, move around the body and may penetrate tissues and remove metals from the inner medium to be accumulated in lysosomes as nondigested products. Rhogocytes also participate in metal mobilization, accumulation, and release. The assessment of metal levels in target cells of sentinel molluscs by microscopic techniques provides an early-warning measure, with promising applications as an exposure biomarker for environmental monitoring programs. Microsc. Res. Tech. 56:358–392, 2002. © 2002 Wiley-Liss, Inc.

SUMMARY Carboxypeptidase E (CPE) is highly concentrated in neuroendocrine tissues and is the only carboxypeptidase detected in mature secretory vesicles. Carboxypeptidase D (CPD), a carboxypeptidase with CPE-like activity, is widely distributed in tissues and is present in the trans -Golgi network. Previous work had shown that both CPE and CPD are expressed in the human placenta and that CPD is

SUMMARY Semicarbazide-sensitive amine oxidase (SSAO), widely distributed in highly vascularized mammalian tissues, metabolizes endogenous and xenobiotic aromatic and aliphatic monoamines. To assess whether its physiological role in humans is restricted to oxidation, we used an immunohistochemical approach to examine the cellular localization of SSAO in human peripheral tissues (adrenal gland, duodenum, heart, kidney, lung, liver, pancreas, spleen, thyroid gland, and blood vessels) and also analyzed its subcellular localization. The results are in agreement with the specific activities also determined in the same samples and are discussed with reference to the tissue distribution of monoamine oxidase A and B. Together with the oxidative deamination of monoamines, SSAO cellular localization indicates that, in most human peripheral tissues, it might participate in the regulation of physiological processes via H2O2generation. (J Histochem Cytochem 49:209–217, 2001)