Phosphotungstic Acid Hematoxylin Staining Research Articles

Finlepsin-retard in clinic of borderline states : V. Ya. Semke, M.M. Aksyonov, I.E. Kupriyanova, A.V. Semke. Mental Health Research Institute, Sosnovy bor, Tomsk, 6340146, Russia

Epistaxis is an important emergency that can sometimes be life threatening without effective intervention. Persistent and recurrent bleeding can lead to aspiration, hypotension, hypoxia, or even severe and mortal cardiovascular complications. Providing prompt hemostasis is important, and the hemostatic method used must be easily and locally applicable, efficient, and inexpensive.The aim of this study was to assess the hemostatic efficacy of Ankaferd Blood Stopper (ABS) in an experimental epistaxis model and to determine the histopathologic alterations with topical ABS application.Twenty-eight New Zealand rabbits were evaluated in 4 study groups. Topical ABS, gelatin foam (GF), adrenalin + lidocaine (AL), and serum physiologic as negative control (C) were applied to the animals for controlling epistaxis. The bleeding was generated with a standard mucosal incision in all groups. Cotton pieces soaked with ABS, AL, C, and GF were applied to the nasal bleeding area. Time of hemostasis was recorded. Tissue samples were obtained after hemostasis for histopathologic examination. The samples were stained with hematoxylin and eosin (HE) and phosphotungstic acid hematoxylin (PTAH) and were examined under a light microscope. In this experimental study, the observers were blind to ABS, AL, and C but not to GF, because of its solid nature.Median durations required for hemostasis in ABS, AL, GF, and C groups were recorded as 30, 90, 90, and 210 seconds, respectively. The time until termination of bleeding in the ABS group was significantly shorter than that in the AL, GF, and C groups (P = 0.002, P = 0.002, and P = 0.001, respectively). On histopathologic evaluation, after staining with HE, minimal fibrin at the incision edges and a few extravasated erythrocytes were observed in the C, AL, and GF groups. In the ABS group, a dark amorphous material surrounded by fibrin, filling the space between the edges of incisions, was noticed. Fibrin was determined in the C, GF, and AL groups with PTAH stain and in the positive control group. In the ABS group, it was observed that the amorphous substance surrounded by fibrin seen in the HE sections was not stained with PTAH.Topical nasal ABS application controlled epistaxis faster than C, GF, and AL in this animal bleeding model. The bleeding model used here might fail to replicate the type of injury that would be likely to result in life-threatening bleeding in humans, which should be considered a limitation of the present study. The histopathologic findings in the nasal incision area suggest that ABS might affect global hemostasis by inducing a unique protein network formation, potentially representing a different mechanism of action among conventional antihemorrhagic applications.

Ultrastructural correlates in the cerebellum of the biochemical effects of lithium on gene expression : S. Sidiropoulou-Skokou, S. Havaki and M.R. Issidorides. Neurobiology Research Institute, Theodor Theohari Cozzika Foundation, and Department of Psychiatry, University of Athens, Eginition Hospital, Greece

Epistaxis is an important emergency that can sometimes be life threatening without effective intervention. Persistent and recurrent bleeding can lead to aspiration, hypotension, hypoxia, or even severe and mortal cardiovascular complications. Providing prompt hemostasis is important, and the hemostatic method used must be easily and locally applicable, efficient, and inexpensive.The aim of this study was to assess the hemostatic efficacy of Ankaferd Blood Stopper (ABS) in an experimental epistaxis model and to determine the histopathologic alterations with topical ABS application.Twenty-eight New Zealand rabbits were evaluated in 4 study groups. Topical ABS, gelatin foam (GF), adrenalin + lidocaine (AL), and serum physiologic as negative control (C) were applied to the animals for controlling epistaxis. The bleeding was generated with a standard mucosal incision in all groups. Cotton pieces soaked with ABS, AL, C, and GF were applied to the nasal bleeding area. Time of hemostasis was recorded. Tissue samples were obtained after hemostasis for histopathologic examination. The samples were stained with hematoxylin and eosin (HE) and phosphotungstic acid hematoxylin (PTAH) and were examined under a light microscope. In this experimental study, the observers were blind to ABS, AL, and C but not to GF, because of its solid nature.Median durations required for hemostasis in ABS, AL, GF, and C groups were recorded as 30, 90, 90, and 210 seconds, respectively. The time until termination of bleeding in the ABS group was significantly shorter than that in the AL, GF, and C groups (P = 0.002, P = 0.002, and P = 0.001, respectively). On histopathologic evaluation, after staining with HE, minimal fibrin at the incision edges and a few extravasated erythrocytes were observed in the C, AL, and GF groups. In the ABS group, a dark amorphous material surrounded by fibrin, filling the space between the edges of incisions, was noticed. Fibrin was determined in the C, GF, and AL groups with PTAH stain and in the positive control group. In the ABS group, it was observed that the amorphous substance surrounded by fibrin seen in the HE sections was not stained with PTAH.Topical nasal ABS application controlled epistaxis faster than C, GF, and AL in this animal bleeding model. The bleeding model used here might fail to replicate the type of injury that would be likely to result in life-threatening bleeding in humans, which should be considered a limitation of the present study. The histopathologic findings in the nasal incision area suggest that ABS might affect global hemostasis by inducing a unique protein network formation, potentially representing a different mechanism of action among conventional antihemorrhagic applications.

Role of Na(+)-H+ exchange on reperfusion related myocardial injury and arrhythmias in an open-chest swine model.

The inhibition of Na(+)-H+ exchange (NHE) with amiloride analogues in vitro has been shown to prevent reperfusion arrhythmias and additional cell necrosis. Inhibition of intracellular Ca2+ overload via NHE inhibition has been suggested as a mechanism of these protective effects. The aim of this study was to examine whether treatment with amiloride analogues reduces the incidence of reperfusion arrhythmias and limits infarct size in vivo. Open-chest swine were exposed to a 30-minute left anterior descending artery (LAD) occlusion and 180 minutes of reperfusion during atrial pacing at 150 ppm. Intravenous 5-(N,N-dimethyl)-amiloride (AML, 5 micrograms/kg per min) was administered in the treatment group (n = 7) and intravenous saline in the control group (n = 7), starting 10 minutes before coronary occlusion. The infusion was continued during ischemia and reperfusion. The area at risk was defined by monastral blue dye and infarct size by triphenyltetrazolium chloride staining. Limb leads ECG and monophasic action potentials (MAPs) from the epicardium in the ischemic area were recorded. There was no significant difference in the size of the area at risk and hemodynamic parameters between the groups. However, the infarcted area was 0.4% +/- 1.0% of the area at risk in the treatment group, whereas it was 62% +/- 29% in the control group (P < 0.05). Pathological examination (Hematoxylin-eosin and Mallory's phosphotungstic acid-hematoxylin staining) revealed that all of the infarcted area consisted of contraction band necrosis. MAP duration in both groups was significantly shortened during ischemia. After reperfusion, MAP duration in the treatment group recovered earlier than that of control group. However, there was no significant difference in the incidence of ventricular tachyarrhythmia between the groups. Inhibition of NHE with AML prevented reperfusion related cell necrosis in the in vivo swine model, but did not reduce the incidence of ventricular tachyarrhythmia.

Juvenile Rhabdomyosarcomas in Two Dogs

Juvenile rhabdomyosarcomas were diagnosed in two young dogs based on the results of histopathology, phosphotungstic acid-hematoxylin stain, immunohistochemistry, and the age of the dogs. One dog, an 11-month-old Rottweiler, had tumor masses in the maxillary gingiva and the urinary bladder. Histologically, the gingival mass was an alveolar type of rhabdomyosarcoma and the urinary bladder mass was an embryonal type. The other dog, a 1.5-year-old Basset Hound, had an embryonal rhabdomyosarcoma involving the oropharynx and the temporal muscles, with metastasis to the regional lymph node and lungs.

Intracytoplasmic eosinophilic hyaline globules in cartilaginous neoplasms: A surgical, pathological, ultrastructural, and electron probe x-ray microanalytic study

Hyaline globules (HGs), spherical intracytoplasmic eosinophilic droplets, have been associated with a variety of conditions, including hepatocellular carcinoma, lung adenocarcinoma, and Kaposi's sarcoma, but they have not been described in cartilaginous tumors. In specimens of 60 cartilaginous neoplasms we found that 22 of 33 chondrosarcomas (67%), eight of 16 enchondromas (50%), and three of seven soft tissue chondromas (43%) exhibited HGs. HGs were seen more commonly in low grade chondrosarcoma (70%) and were relatively rare in high grade chondrosarcoma (25%). No HGs were identified in three osteochondromas, one synovial chondromatosis, or 15 normal cartilaginous tissues taken from various sites. Cartilage associated HGs ranged in size from 2 to 20 μm, were diastase resistant and periodic acid-Schiff (PAS) stain positive, demonstrated autofluorescence, and variably stained with Mallory's phosphotungstic acid-hematoxylin stain (PTAH). A panel of immunostains did not show any specific staining reactions with HGs. Ultrastructurally the HGs were spherical, non-membrane-bound bodies having complex architectural features associated with profiles of rough endoplasmic reticulum. Electron probe x-ray microanalytic (EPXMA) study showed significant peaks of sulphur and calcium. We conclude that HGs represent secretory products of probable glycoprotein nature, may accumulate in a variety of cartilaginous neoplasms, and may be seen more frequently in low grade chondrosarcomas.

Fibrin content of carotid thrombi alters the production of embolic stroke in the rat.

Mechanical denudation of the endothelium of the carotid artery in animals produces a nonocclusive thrombus, but the brains of these animals have not been examined for the presence of embolic stroke. The endothelium of the right carotid artery of 16 Wistar rats was denuded using a balloon catheter. Phosphotungstic acid hematoxylin (PTAH) staining and scanning electron micrographs of the nonocclusive thrombi in the carotid arteries were compared with those produced by photochemical methods, and brains were examined for infarcts. Although nonocclusive thrombi were present in the carotid arteries of 4 of 4 rats killed at 4 hours and in 8 of 12 killed at 24 hours, neither cerebral infarcts nor emboli were seen in the 14 brains evaluated by light microscopy. PTAH demonstrated a high fibrin content in the thrombus produced by the endothelial denudation, with almost no fibrin seen in photochemically induced thrombi. Scanning electron microscopy confirmed dense networks of fibrin in the thrombi produced by balloon denudation. The composition of a nonocclusive thrombus may determine the embolic potential of this thrombus. A low fibrin content in a nonocclusive platelet thrombus may enhance the embolic potential. This suggests that platelet inhibition may also be indicated in patients with carotid artery disease who are being treated with anticoagulant.

Rapid Microwave Phosphotungstic Acid-Hematoxylin Stain for Paraffin and Glycol Methacrylate Sections

AbstractA microwave modification of the Mallory phosphotungstic acid hematoxylin stain is detailed. The technique eliminates the use of lengthy toxic mercuric chloride mordanting and dezenkerization by substituting zinc chloride. Staining is carried out at low heat levels to preclude boiling and stain breakdown and results in sharp, contaminant-free, dark blue muscle striations within about 20min in paraffin and glycol methacrylate tissue sections. (The J Histotechnol 17:125, 1994)

Relationship of clear cell oncocytoma to mitochondrial-rich (typical) oncocytomas of parotid salivary gland: An ultrastructural study

Nosologically, the designation clear cell oncocytoma seems illogical and contradictory when the definition of an oncocytoma relies on mitochondrial-rich cells; its relationship to the granular (mitochondrial-rich) oncocytoma needs further clarification. Electron microscopy of six examples of oncocytoma from five patients allowed assessment of cellular features in three clear cell variants, two typical oncocytomas and one with a roughly equal proportion of clear and granular tumor cells. Ultrastructurally, in the clear cell types of oncocytoma, a considerable to extensive portion of the central cytoplasm was occupied by monoparticulate glycogen with margination of mitochondria and other organelles. The degree of extraction of the glycogen during fixation and processing accounts for variation in the extent of the clear cell component as phosphotungstic acid-hematoxylin staining reveals considerable mitochondria in many of the tumor cells in the clear cell variant of oncocytoma. Ultrastructural findings further support an interrelationship between clear and granular cells, as some typical oncocytes are evident in the clear cell variant of oncocytoma, and one oncocytoma in this series had a roughly equal number of glycogen-rich and mitochondrial-rich tumor cells both of which took part in the formation of microlumens. The designation clear cell oncocytoma is appropriate within the oncocytoma subgroup of salivary gland tumors.

Rapid Microwave Phosphotungstic Acid-Hematoxylin Stain for Paraffin and Glycol Methacrylate Sections

Rapid Microwave Phosphotungstic Acid-Hematoxylin Stain for Paraffin and Glycol Methacrylate Sections

Morphometrical evaluation of triflusal in brain infarction.

MCA occlusion in animals is a common model for experimental stroke. In previous studies we have shown that one of the factors, which influence evolution of an infarct is microthrombosis in the area of infarction and in the surrounding brain tissue. The present study was undertaken for assessment of the number of microthrombi and of the size of brain infarcting in rats treated with the antiaggregatory substance Triflusal. 7 groups of Sprague-Dawley rats, each group consisting of 6 animals, underwent transsphenoidal MCA occlusion. The animals received Triflusal in various amounts from day 2 till day 6. At day 7 animals were decapitated and the brains were fixed in formaldehyde. The brain was dissected at the level of the optic chiasm and embedded in paraffin. Fresh microthrombi were detected py PTAH (Phosphotungstic acid hematoxylin) staining. In each animal the hemisphere with the ischemic lesion as well as the contralateral hemisphere were examined. The area of both hemispheres was calculated by subtraction of the ventricle area from the total brain area of a section. Infarct was defined as the region of necrosis which was sharply demarcated from normal brain. The infarcted area was planimetrically measured to obtain a ratio of infarcted to normal brain. A correlation between the effect of Triflusal, number of microthrombi and size of the infarcted area could be demonstrated. The pathogenetic role of the microthrombi in the evolution of cerebral infarction as well as the effect of Triflusal in different dosages on the number of microthrombi could be clearly assessed by quantitative morphometry.

Hair Follicle Nevus with Hyperplasia of Smooth and Striated Muscle

We report a peculiar nevus which occurred on the left side of the chin of an 18-year-old male. It had been present for four years and had grown rapidly during the last year. Histologically, many hair follicles, smooth muscles, and striated muscles were found within the reticular dermis. The hair follicles were accompanied by mature sebaceous glands; numerous eccrine glands were seen in the middle and deep dermis. Immunohistochemical stain and phosphotungstic acid hematoxylin stain supported the fact that the muscles seen between the follicles were both smooth and striated. This nevus was considered to be a hair follicle nevus accompanied by hyperplasia of smooth muscles and striated muscles.

&lt;I&gt;Infantile Myofibromatosis with a Solitary Lesion in the Skull&lt;/I&gt;

The term "infantile myofibromatosis" was coined in 1981 to describe a rare type of soft-tissue tumor in infants. Solitary lesions are usual, but extremely rare in the skull. An infantile case involving a lesion in the left parietal bone is described. The patient was a 6-month-old girl admitted with a mass measuring 2 x 2 cm in the left parietal region. Skull x-rays showed an osteolytic lesion with a sclerotic margin in the parietal bone. Computed tomographic (CT) scans revealed a low-density mass with homogeneous enhancement. The tumor adhered to the dura and had destroyed the left parietal bone. Histological examination disclosed spindle-shaped cells arranged in short bundles and abundant vasculature. Phosphotungstic acid hematoxylin staining revealed longitudinal fibrils resembling myofibroblasts. This is the first report in which CT findings are described in a case of infantile myofibromatosis with a solitary lesion occurring in the skull.

Congenital Mesoblastic Nephroma in a Dog: A Benign Variant of Nephroblastoma

A nephroblastoma consisting predominantly of mature mesenchymal components developed in a 7-month-old male beagle dog. The dog was necropsied after experimental use. There were no abnormal signs or laboratory tests prior to necropsy. The left kidney was slightly enlarged, and the tumorous area of the kidney was gray-white. The cut surface of the tumor, occupying the upper one-third of the kidney, revealed a solid, rubbery, yellow-white mass. The tumor was about 3.0 cm in its greatest diameter, showed replacement growth pattern, and lacked a capsule (Fig. 1). Histologically, the tumor was mainly composed of spindle cells arranged in interlacing bundles that were intermingled with the normal renal tissue; however, there was no sharp demarcation between bundles and the surrounding renal parenchyma. The greater part of the tumor had a loose myxomatous pattern with an alcian blue-positive matrix, but dense, cellular, fibromatous areas with transition to the myxomatous areas were also noted (Fig. 2). Spindle cells (fibroblasts) had scanty cytoplasm with cigar-shaped nuclei and were negative for phosphotungstic acid-hematoxylin staining. Neither nuclear atypia nor mitotic figures were detected. Within the myxomatous area, cysts of various sizes were recognized in addition to entrapped nephrons. The small cysts were dilated

Mucin-producing bronchioalveolar-cell carcinoma. With special reference to a characteristic structure revealed by phosphotungstic acid-hematoxylin staining

Mucin-producing bronchioalveolar-cell carcinoma. With special reference to a characteristic structure revealed by phosphotungstic acid-hematoxylin staining

Immunohistochemical demonstration of glial fibrillary acidic protein (GFAP) in nasal gliomas

Using the Sternberger method (Immunoluk Histoset KIT) GFAP (glial fibrillary acidic protein) was demonstrated immunohistochemically in 4 nasal gliomas. In these histologically complex tumour-like lesions mesenchymal, epithelial, and neuroglial tissues as well as small groups of scattered glial elements could be differentiated specifically by the highly sensitive GFAP immunoperoxidase technique. GFAP was present in astrocytes and astrocyte-like differentiations. The reactivity of cell processes was essentially lower. The GFAP immunostain does not always correlate with Mallory's phosphotungstic acid hematoxylin (PTAH) stain and Gallyas' silver impregnation method for astrocytes. Additionally the immunohistochemical investigation of semithin sections prepared by the so-called pop off technique after Bretschneider et al. (1981) allows the correct localization of GFAP in astrocytes and their modulations. Furthermore, in this study, the intimate connection of epithelium and glial cells as well as astrocytes containing hemosiderin granules could be demonstrated. The latter findings suggest a possible phagocytotic activity of astrocytes. Our results show that the demonstration of GFAP by the Sternberger method is a valuable aid in establishing astrocytic glial differentiations and modulations in complex tumour-like lesions such as nasal gliomas.

DISSEMINATED INTRAVASGULAR COAGULATION (DIC)

Renal tissues of 208 autopsied cases were examined. Malignant neoplasm with hematological malignancies often accompanied DIC. Tissue sections were stained with hematoxylin and eosin and Mallory's phosphotungstic acid hematoxylin (PTAH), and were applied for immunoperoxidase method (IP), using antisera against human fibrinogen, FDP-D and FDP-E. Histologically in 80 cases (38%) fibrin or fibrinogen related materials (FRMs) were observed in the glomerular capillary or the intratubular area or in both. FRMs were PTAH or IP positive or both in 23 of the 26 cases (88%) clinically diagnosed as DIC. In the remaining three cases anticoagulants probably interfered with FRMs observation. This study showed the PTAH stain was nonspecific and insensitive to FRMs, and that IP was necessary for a pathological diagnosis of DIC. The presence of FRMs in the renal tubuli is an important finding in confirming DIC. DIC may be present histologically in the absence of clinical DIC symptoms.

Infantile digital fibromatosis after web construction in syndactyly

A case of infantile digital fibromatosis with an unusual onset is reported. A Japanese girl, with a simple syndactyly of the right ring and little fingers, had an operation at the age of 2 1/2 years. Three months after the operation, multiple nodules appeared at the skin graft edge. The nodules were excised but soon recurred. When the patient was 8 years old, a large-scale excision of the tumors was performed. Intracytoplasmic inclusion bodies were shown by phosphotungstic acid-hematoxylin stain, and the case was diagnosed as infantile digital fibromatosis.

Isolation and properties of rat cell lines morphologically intermediate between cultured mammary epithelial and myoepithelial-like cells

The cloned cuboidal epithelial cell line Rat Mammary (Rama) 25 converts at low frequency in culture to elongated cells that possess some of the properties of myoepithelial cells; one such clonal cell line is termed Rama 29. Three morphologically intermediate clonal cell lines have been isolated from Rama 25 which form a morphological series in the order: Rama 25 cuboidal cells, Rama 25-Intermediate 2(I2), Rama 25-I1, Rama 25-I4, and Rama 29 elongated cells. This same order is largely maintained for increasing percentages of elongated cells, decreasing percentages of cuboidal cells, decreasing tubular structures on collagen gels, and increasing times of appearance of tumors in nude mice. The fully elongated cells fail to revert to cuboidal cells and to form tumors. Binding of antisera to epithelial-specific milk fat globule membranes and human keratin declines whereas binding of antisera to myoepithelial-associated laminin, vimentin, and Thy-1 increases in the cell lines in the same order. Similarly 7 polypeptides characteristic of elongated cells increase and 4 polypeptides characteristic of cuboidal cells decrease in the cell lines in the same way. Anti-actin serum binds equally to all cell lines grown on plastic, except for Rama 25-I4, where its binding is increased. Rama 25-I1 and Rama 25-I4 cells also give rise to anti-actin, anti-myoglobin, and phosphotungstic acid hematoxylin-staining giant, striated cells on collagen gels and in tumors that also have ultrastructural characteristics of skeletal muscle. Fresh elongated converts of Rama 25 bind appreciably more anti-actin serum than many of the clonal elongated cell lines such as Rama 29. Ultrastructural analysis confirms the gradual loss of epithelial characteristics and the acquisition of immature myoepithelial characteristics in the same sequence of cell lines. It is suggested that such a linear sequence of intermediate morphological states occurs between the Rama 25 cuboidal cells and the elongated myoepithelial-like cells in vitro, and that a similar morphological sequence may exist in terminal ductal structures in vivo.

Functioning oxyphil cell adenoma of the parathyroid gland: A clinicopathologic study of ten patients with hyperparathyroidism

Adenomas of the parathyroid gland, the majority of which are of the solitary chief cell type, are the most frequent cause of primary hyperparathyroidism. Parathyroid adenomas composed predominantly or exclusively of oxyphil cells are rare and were previously considered nonfunctioning. Among 160 consecutive patients with primary hyperparathyroidism who were explored surgically, ten (6.25 per cent) had adenomas of the oxyphil cell variety. The eight women and two men in this series ranged in age from 28 to 82 years (average, 58 years). The tumors weighed from 0.2 to 4.0 g (average, 1.2 g). Nine of the adenomas were located in the left neck, while the remaining tumor was in the right neck. Histologically, the tumors were composed almost exclusively of transitional and typical oxyphil cells. A residual "capsular" parathyroid composed of chief cells was identified in each case. Electron microscopy, phosphotungstic acid-hematoxylin staining, and the immunoperoxidase technique for cytochrome c oxidase showed the presence of numerous mitochondria in oxyphil cells. All patients had normal serum calcium levels four months to four years (average, 23 months) after removal of the neoplasms.

Establishment and Characterization of Two Human Mixed Mesodermal Tumor Cell Lines From the Same Patient

A cell line designated HIRS -BM was established from fluid aspirated from the sternal bone marrow of a 16-year-old female. Another cell line ( HIRS -PB) was derived from the peripheral blood of the same patient. Both lines grew well, multilayering rapidly without contact inhibition, and 62 serial passages were successively done within 28 months. Both cultures contained spindle- or fibrous-shaped cells that revealed neoplastic and pleomorphic features, and these cells were characterized as possessing cross-striations in the cytoplasm. The cross-striations were detected by phosphotungstic acid hematoxylin stain. Some elongated cells were stained positively with anti-myoglobin by use of periodic acid-Schiff methods. The primary tumor in the uterus was diagnosed as a mixed mesodermal tumor composed of adenocarcinoma and rhabdomyosarcoma cells. The karyotype exhibited hyperploidy and large submetacentric marker chromosomes, and the modal chromosome number was 84. No difference was found between the 2 cell lines except for growth behavior and heterotransplantability . HIRS -BM cells grew more rapidly and were highly transplantable. The HIRS -BM cells were transplanted into the subcutis of BALB/c nude mice and produced mixed mesodermal tumors resembling the uterine tumor, while the HIRS -PB cells could not be transplanted. Due to the histogenesis of the mixed mesodermal tumor being's obscure with histologic observations only, this study was performed to obtain data by tissue culture of the tumor and resulted in support of the combination theory reported in the literature in regard to tumor.