The peritubular myofibroblasts in the testes from normal men and men with Klinefelter's syndrome. A quantitative, ultrastructural, and immunohistochemical study.

The ultrastructure and immunostaining with antibodies against actin, desmin, and vimentin were studied in the peritubular myofibroblasts of testes from normal men and men with Klinefelter' syndrome (KS). The seminiferous tubules were classified into five types (a-e), related to the progressive degree of sclerosis measured as thickening of the lamina propria. In control testes, only types a and b tubules were present, whereas the testes from men with KS showed types b, c, d, and e tubules. The ultrastructural study revealed abundant microfilament bundles with electron-dense bodies in the cell periphery of the myofibroblasts in a and b tubules. In c tubules, the microfilament bundles of the myofibroblasts were lacking in electron-dense bodies. Myofibroblasts in tubules d and e showed scanty microfilament bundles. Immunostaining of peritubular myofibroblasts with anti-actin antibodies was intense in tubule types a-c and scanty in types d and e. Immunostaining of myofibroblasts with anti-desmin antibodies was intense in tubule types a and b, and negative in types c-e. Immunostaining with anti-vimentin antibodies was weak in tubule types a-c and intense in types d and e. Quantitative study revealed that with the progression of sclerosis, the number and volume per cross-sectioned tubule of actin-containing cells and, mainly, desmin-containing cells decrease while the number and volume of vimentin-containing cells increase.     

Integrin pattern and effect on contraction in cultured testicular peritubular myoid cells

PROBLEM: Neither the integrin pattern nor the biological functions of integrins have been extensively documented in human cultured testicular peritubular myoid cells (TPMC). The integrin pattern and the presence of some proteins of the immunoglobulin superfamily on human TPMC as well as the role of integrins in TPMC contraction were examined. METHOD OF STUDY: Integrin expression was evaluated by immunofluorescence and FACS analysis. To assess the role of integrin in TPMC contraction, human and rat cells were added to a collagen gel system and exposed to contractile stimuli. RESULTS: The immunofluorescence and cytofluorimetric analysis showed that human cultured TPMC express alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, alphav, beta1, beta3, and beta4 integrin subunits, and significant amounts of intercellular adhesion molecule-1 (ICAM-1), whereas they do not present alpha4, beta2, beta7 subunits, nor intercellular adhesion molecule-2 (ICAM-2) and neural cell adhesion molecule (NCAM). The preincubation of human cells with an anti-beta1 mAb and of rat cells with a polyclonal anti-beta1 antibody inhibited TPMC contraction induced by different contractile stimuli. CONCLUSION: Our investigation documented a broad integrin pattern on human cultured TPMC as well as a role for integrins in human and rat TPMC contraction.     

Identification of smooth muscle myosin in myoid cells of the testes

Smooth muscle myosin was found by the Coons' immunomorphological test in the outer wall of the seminiferous tubules of man, rats, and mice. The results of the investigation confirm the smooth-muscle nature the myoid cells.Laboratory of General Pathological Anatomy, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. I. Strukov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsniny, Vol. 82, No. 12, pp. 1499–1501, December, 1976.     

茶多酚对精索静脉曲张大鼠生精细胞凋亡的影响及机制研究

目的探究茶多酚(TP)对精索静脉曲张(Vc)SD大鼠生精细胞凋亡及细胞色素c(Cyt-c)、波形蛋白表达的影响及机制。方法将36只SD大鼠分为对照组、Vc组和Vc+TP组,每组12只。Vc组和Vc+TP组通过左肾静脉结扎构建精索静脉曲张模型,Vc+TP组大鼠给予茶多酚(40 mg/kg)灌胃,对照组和Vc组以等量生理盐水灌胃,连续4周。干预4周后取出大鼠睾丸组织,通过HE染色和TUNEL染色观察睾丸组织损伤和生精细胞凋亡情况,并检测茶多酚对氧化应激指标的影响。通过RT-qPCR和Western blot检测Cyt-c和波形蛋白表达。结果HE染色结果显示,Vc组出现广泛的上皮损伤,Vc+TP组损伤情况明显轻于Vc组。Vc组凋亡指数高于对照组及Vc+TP组(P<0.05)。Vc组丙二醛(MDA)水平高于对照组及Vc+TP组(P<0.05),而超氧化物歧化酶(SOD)水平低于对照组及Vc+TP组(P<0.05);Vc组波形蛋白mRNA和蛋白表达水平均低于对照组及Vc+TP组(P<0.05),而Cyt-c mRNA和蛋白表达水平均高于对照组及Vc+TP组(P<0.05)。结论茶多酚可通过抑制氧化应激反应保护线粒体功能并促进波形蛋白的表达,从而抑制生精细胞凋亡,减轻睾丸损伤,达到缓解精索静脉曲张的目的。     

Elevation of serum testosterone and free testosterone after embolization of the internal spermatic vein for the treatment of varicocele in infertile men

BACKGROUND: To evaluate the effect of internal spermatic vein (ISV) embolization on levels of serum testosterone and free testosterone and on spermatogenesis. METHODS: The files of 83 infertile men treated for varicocele were reviewed for changes in serum testosterone, free testosterone and spermatogenesis after ISV embolization. RESULTS: Mean serum testosterone concentration rose after embolization by 43%, from 12.07 +/- 6.07 nmol/l to 17.22 +/- 8.43 nmol/l (P<0.001). Mean serum free testosterone concentration rose by 72%, from 5.93 +/- 2.44 nmol/l to 10.21 +/- 7.69 nmol/l (P<0.001). Mean sperm concentration increased from 7.49 +/- 1.73 x 10(6)/ml to 18.14 +/- 2.36 x 10(6)/ml (P<0.001); mean sperm motility increased from 21.74 +/- 2.47 to 34.47 +/- 2.27% (P<0.001); and mean sperm morphology increased from 6.63 +/- 1.07 to 13.08 +/- 1.44% (P<0.001). CONCLUSIONS: ISV embolization apparently induces an increase in both serum testosterone and free testosterone concentrations and in sperm parameters in infertile patient with varicocele, regardless of the size of the varicocele.     

Fibrous hamartoma of infancy: a histochemical and immunohistochemical study

Fibrous hamartoma of infancy is an uncommon lesion, the histogenesis and biological nature of which are uncertain. Ten cases have been studied by light microscopy, mucin histochemistry and immunohistochemistry. The typical histological features are presented. The presence of hyaluronic acid and of chondroitin-4- and -6-sulphate and keratan sulphate has been demonstrated in different components of the tumour. Vimentin positivity was noted in the undifferentiated and fibroblastic components. The implications of these findings are discussed. Whether the lesion merits the designation of hamartoma or is, in fact, a benign neoplasm remains obscure.     

The study of varicocele through the use of animal models

The pathophysiology of the varicocele has received considerable study, both in humans and in animal models. Mechanistic information is difficult to obtain from human subjects because study designs must not be invasive and the subject population is variable in the status of the varicocele, patient age, fertility or other health-related issues. Because of these limitations, animal models of varicocele have been developed in several species, the most common being the rat. Surgery to establish the varicocele involves partial obstruction of the left renal vein, causing a varicosity of the left spermatic vein, including the pampiniform plexus. Studies using this model have shown that experimental left varicocele induces bilateral increases in testicular blood flow and temperature contemporaneous with decreases in intratesticular testosterone and testicular sperm output. Spermatic vein reflux is not related to the pathophysiological consequences of experimental varicocele. Many questions remain regarding the mechanism by which varicocele induces testicular dysfunction, chief among them being how the unilateral varicocele causes a bilateral testicular response in the first place.     

Fusocellular gonadal stromal tumour of the testis with epithelial and myoid differentiation

We describe an unusual fusocellular gonadal stromal tumour with a benign behaviour in the left testis from a 16-year-old man. The neoplasm consisted of a non-encapsulated proliferation of irregularly arranged, fusiform cell bundles in fibrous connective tissue. The tumour cells contained a slightly infolded nucleus, some dilated rough endoplasmic reticulum cisternae, abundant filament bundles which connected to subplasmalemmal electron-dense bodies, pinocytotic vesicles and a discontinuous basal lamina. The intercellular spaces were narrow and the tumour cells were joined by desmosomes. These cells were immunoreactive for muscle actin, α-actinin and vimentin. Focal immunostaining for collagen type IV was observed around the cells. No immunoreactivity for keratins, desmin S-100 protein or XIIIa factor was found. The findings suggest that the tumour arose from the peritubular myoid cells.     

Gastrointestinal stromal tumor of the stomach with rhabdoid phenotype: immunohistochemical, ultrastructural, and immunoelectron microscopic evaluation

A variety of neoplasms with rhabdoid differentiation have been reported in many sites. The authors describe a case of gastrointestinal stromal tumor (GIST) of the stomach that exhibited prominent rhabdoid features. Immunohistochemically, the tumor cells displayed positive staining for vimentin, c-kit, CD34, and alpha smooth muscle actin. Ultrastructural examination of the rhabdoid tumor cells revealed paranuclear whorls of intermediate filaments, which were immunoreactive for vimentin by both light microscopic immunohistochemical and protein A gold immunoelectron microscopic techniques. On H&E light microscopic examination alone, such a tumor could be mistaken for a variety of epithelial, mesenchymal, or other neoplasms that may show rhabdoid features. One report of GIST with a rhabdoid histologic phenotype has been described. This is the second known report of such a case with immunophenotypic and ultrastructural evaluation, and the first case with immunoelectron microscopic examination.     

Human decidual stromal cells express alpha-smooth muscle actin and show ultrastructural similarities with myofibroblasts.

Previous reports in human and mouse material demonstrated that decidual stromal cells expressed antigens associated with haematopoietic cells, exerted immune functions, and originated from bone marrow. These findings suggested that these cells belonged to the haematopoietic lineage. We purified and expanded in culture precursors of human decidual stromal cells, and found in electron microscopic images that the ultrastructure of these cells was similar to that of myofibroblasts, which are of mesenchymal origin. The relationship between these two types of cell was confirmed by the detection (by flow cytometry) in the decidual precursors of alpha-smooth muscle actin, a contractile microfilament expressed solely by smooth muscle cells, myofibroblasts and related cells. This filament was also detected in decidual stromal cells decidualized in vitro by the effect of progesterone. We also found vimentin in decidual precursors and decidualized cells. This intermediate filament has been previously reported to be expressed by all decidual stromal cells and also by myofibroblasts. Desmin, another intermediate filament expressed by myofibroblasts, was not detected in the decidual precursors; however, this filament was observed in decidualized cells. The expression of alpha-smooth muscle actin by decidual stromal cells was also found by immunostaining in cryostat sections of early decidua. Our results suggest that decidual stromal cells are related to myofibroblasts.     

Apical mitochondria-rich cells in the human epididymis: an ultrastructural, enzymohistochemical, and immunohistochemical study.

An ultrastructural, enzymohistochemical, and immunohistochemical study of the ductus epididymis in normal men was undertaken to investigate the characteristics of the apical mitochondria-rich cells (AMRCs). These cells, which differ morphologically from the principal cells (PCs), appear in isolation in the caput epididymidis (5.8 +/- 1.7 cells per cross-sectional duct) and only occasionally in the corpus epididymidis. The morphologic appearance of AMRCs varies from slender cells extending from the basement membrane to the lumen to apical cells without apparent contact with the basement membrane. The former display a round pale nucleus located in the middle of the epithelium; the apical cells have a dark nucleus, which, surrounded by a narrow cytoplasmic band, protrudes into the lumen. The cytoplasm of AMRCs is electron-dense and contains numerous mitochondria surrounded by rough endoplasmic reticulum cisternae. In the apical portion, there are lysosomes, vesicles with an electron-dense granule, and vacuoles showing a variable size and content. The stereocilia are shorter and less numerous than those of the PCs. The AMRCs are similar to the PCs in the intensely positive reaction for the enzymatic activity acid phosphatase, as well as in the lack of reaction for alkaline phosphatase and phosphorylase activities. AMRCs differ from PCs in: (1) a more intense reaction to the enzymatic activities ATPase, NADP, and succinic dehydrogenease, (2) a more intense immunostaining by AE1/AE3 and Ks4.62 anti-cytokeratin antibodies, and anti-estradiol receptor protein (D5) antibodies, and (3) a lower staining affinity for epithelial membrane antigen (EMA) antibodies. No positive immunostaining for the anti-cytokeratin Ks8.6 antibodies was observed in either AMRCs or PCs.     

The widespread distribution of Langerhans cells in pathologic tissues: an ultrastructural and immunohistochemical study

Ultrastructural and/or immunohistochemical analysis of approximately 500 specimens revealed widespread distribution of Langerhans cells in pathologic specimens. More than half of the tissue specimens were from patients with a variety of pulmonary diseases. In all specimens in which Langerhans cells were identified ultrastructurally, they were also identified by an immunoperoxidase technique for the visualization of S-100 protein; the latter technique also revealed the dendritic nature of these cells. Langerhans cells were present in 80 to 93 per cent of 66 pulmonary adenocarcinomas and 17 per cent of squamous cell lung cancers; they were not observed in neuroendocrine lung carcinomas or mesotheliomas. They were also observed in benign inflammatory conditions of the lung of several types and in a variety of other malignant neoplasms and disease processes. The Langerhans cells in these tissues varied from few to many. They were most numerous in bronchioloalveolar cell carcinoma, sometimes appearing almost as frequently as tumor cells, and rare in some benign pulmonary conditions and other tumors. The function of Langerhans cells in these conditions is unknown, but they may have an immunologic function, such as antigen processing and presentation to T lymphocytes.     

Intervertebral disc amyloidosis: histochemical, immunohistochemical and ultrastructural observations

Intervertebral discs selected at random from autopsies and surgical operations on herniated discs were examined for evidence of amyloid deposition. Amyloid deposits were detected in 53 (81.5%) of 65 autopsy cases. Discs from subjects over 50 years of age revealed a significantly high incidence of amyloid deposition. Among herniated discs, amyloid deposits were documented in 49 (75.4%) of 65 surgical specimens. The incidence of amyloid deposition in intervertebral discs increased with advancing age. Intervertebral disc amyloidosis consisted of amyloid deposits of three morphological types: linear amyloid deposits around the degenerating chondrocytes (perichondrocyte type), and nodular or band-like deposits in the annulus fibrosus (annulus fibrosus type) and nucleus pulposus (nucleus pulposus type). We suspect that the precursor protein of perichondrocyte type amyloid is derived from chondrocytes, and those of annulus fibrosus type and nucleus pulposus types are derived from the blood. The amyloid deposits were resistant to treatment with potassium permanganate. Immunohistochemically, the amyloid deposits reacted with antibody against amyloid P-component, but not with antibodies against AA, A k , Aλ, transthyretin or β₂-microglobulin. Ultrastructurally, the amyloid deposits were composed of 10 nm-wide nonbranching fibrils. The amyloid in herniated discs had the same biochemical and immunohistochemical properties as those found in autopsy cases. The immunohistochemical characteristics of the amyloid deposits suggest that it derives from an, as yet, unknown precursor protein.     

Mucinous colorectal adenocarcinoma with signet-ring cells: immunohistochemical and ultrastructural study

A primary mucinous colorectal adenocarcinoma tissue with signet-ring cells, as revealed after histological evaluation, was examined ultrastructurally. The authors also analyzed the immunohistochemical data of the tissue for serotonin, vasoactive intestinal polypeptide (VIP), bombesin, somatostatin, and glucagon, using the peroxidase anti-peroxidase (PAP) method and the immunogold labeling method for light and electron microscope, respectively. Electron microscopically mucinous adenocarcinoma was characterized by the formation of small lumen. Adenocarcinoma cells were full of mucous granules of varying electron density, providing a good environment for the tumor cells to grow. They also exhibited a significant loss of microvilli and intracytoplasmic junctions, which could allow the cells to disseminate. Signet-ring cells were located in the basal site of the ducts or in the lamina propria and appeared neoplastic, with mucin accumulation intracellularly and an eccentric crescent-shaped nucleus. The cytoplasmic organelles were decreased and at the periphery of the cell. The PAP method demonstrated that these cells were strongly positive for bombesin and also positive for vasointestinal polypeptide (VIP). The immunogold method detected bombesin immunoreactivity in the vacuoles as well as in other cytoplasmic membranes, whereas VIP was localized mainly in the plasma membrane. The location of signet-ring cells combined with the immunoreactivity for bombesin and VIP indicated that signet-ring cells were of neuroendocrine origin and probably dedifferentiated enterochromaffin-like endocrine cells. These findings have implications for understanding the biological behavior of these composite malignant tumors and could help in the knowledge of the origin of signet-ring cells.     

Immunohistochemical and quantitative study of interstitial and intratubular Leydig cells in normal men, cryptorchidism, and Klinefelter's syndrome.

Testicular specimens from normal men and men with cryptorchidism (CR) or Klinefelter's syndrome (KS) were taken, processed for light microscopy, and stained with the avidin-biotin peroxidase complex method for immunohistochemical detection of testosterone. The Leydig cells were classified by their morphology (normal, multivacuolated, and pleomorphic Leydig cells) and by their staining affinity for anti-testosterone antibodies (T-, T+, and T++ cells), and the average numbers of each cell type for each group of testes were calculated. Normal testes showed morphologically normal interstitial Leydig cells (96.0 +/- 10 per cent) and multivacuolated Leydig cells (4.0 +/- 1 per cent). Cryptorchid testes showed normal Leydig cells (85.8 +/- 11 per cent) and multivacuolated Leydig cells (14.2 +/- 2.3 per cent). Men with KS showed normal Leydig cells (78.9 +/- 9.1 per cent), multivacuolated Leydig cells (9.2 +/- 1.2 per cent), and pleomorphic Leydig cells (11.0 +/- 1.8 per cent). The percentage of T++ cells was higher in normal testes (29.4 +/- 2.1 per cent) than in CR (11.4 +/- 2.2 per cent) and KS testes (6.3 +/- 0.7 per cent). This suggests reduced functional Leydig cell activity in CR and KS. Multivacuolated Leydig cells showed weaker immunostaining than did normal Leydig cells in all the testicular groups. No immunostaining was shown by pleomorphic Leydig cells. Intratubular Leydig cells were only found in CR and KS. Immunostaining was weaker in intratubular Leydig cells than in interstitial Leydig cells. This suggests that intratubular location reduces functional activity of Leydig cells.     

Androgen insensitivity syndrome: an immunohistochemical, ultrastructural, and morphometric study

OBJECTIVE: To evaluate the morphometric, immunohistochemical, and ultrastructural lesions of the testes in prepubertal and adult patients with androgen insensitivity syndrome. METHODS: We examined the testicular biopsy using immunohistochemistry for vimentin, smooth muscle actin, and collagen IV antigens. Quantification of seminiferous tubules and testicular interstitium was performed in prepubertal and adult patients with androgen insensitivity syndrome and results were compared with normal testes from both infants and adults. RESULTS: The adult testes presented nodular and diffuse lesions that consisted of Sertoli-cell-only seminiferous tubules. Two types of Sertoli cells could be distinguished, namely, immature vimentin-positive Sertoli cells and nearly mature Sertoli cells. In the nodules, the lamina propria was thin and contained a scant number of actin-positive peritubular cells. Leydig cells were hyperplastic. The prepubertal patients showed only diffuse lesions characterized by Sertoli cell hyperplasia, decreased germ cell numbers, and a discontinuous immunoreaction to collagen IV. CONCLUSIONS: The testicular lesions in androgen insensitivity syndrome are probably caused by primary alterations that begin during gestation. These lesions become progressively more pronounced at puberty, when the nodular lesion pattern (adenomas) is completely developed.     

Heterogeneity of epithelial cells and reactive components in thymomas: an ultrastructural and immunohistochemical study

Fourteen thymomas were studied by electron microscopy and immunohistochemistry. Based on the ultrastructure of the neoplastic epithelial cells in comparison with normal thymic epithelium, four cortical-, three mixed-, five medullary-, and two corpuscular-type tumors were categorized. Histologically the tumors of cortical type showed prominent lymphocytic infiltration, but scant interdigitating reticulum cells (IDCs) were demonstrated by immunoperoxidase method on paraffin sections with anti-S-100 protein antiserum. Fewer lymphocytes but more IDCs were present in the tumors of medullary and corpuscular types, although variable in those of mixed type. This corticomedullary difference among thymomas was confirmed in some of them by the immunoperoxidase method on frozen sections with monoclonal antibodies. The cortical-type tumors were HLA-DR positive in tumor cells and infiltrated predominantly with cortical thymocytes (OKT-6+, OKT-3-, both Leu 3a/3b+ and OKT-8+), whereas the medullary- and corpuscular-type tumors were HLA-DR positive primarily in IDCs but not in tumor cells and were infiltrated more with medullary thymocytes (OKT-6-, OKT-3+, either Leu 3a/3b+ or OKT-8+). The classification of thymomas based on neoplastic epithelial cells will serve to refine the traditional classification based on reactive lymphocytes.     

Apical mitochondria-rich cells in the human epididymis: An ultrastructural,enzymohistochemical, and immunohistochemical study

An ultrastructural, enzymohistochemical, and immunohistochemical study of the ductus epididymis in normal men was undertaken to investigate the characteristics of the apical mitochondria-rich cells (AMRCs). These cells, which differ morphologically from the principal cells (PCs), appear in isolation in the caput epididymidis (5.8 ± 1.7 cells per cross-sectional duct) and only occasionally in the corpus epididymidis. The morphologic appearance of AMRCs varies from slender cells extending from the basement membrane to the lumen to apical cells without apparent contact with the basement membrane. The former display a round pale nucleus located in the middle of the epithelium; the apical cells have a dark nucleus, which, surrounded by a narrow cytoplasmic band, protrudes into the lumen. The cytoplasm of AMRCs is electron-dense and contains numerous mitochondria surrounded by rough endoplasmic reticulum cisternae. In the apical portion, there are lysosomes, vesicles with an electron-dense granule, and vacuoles showing a variable size and content. The stereocilia are shorter and less numerous than those of the PCs. The AMRCs are similar to the PCs in the intensely positive reaction for the enzymatic activity acid phosphatase, as well as in the lack of reaction for alkaline phosphatase and phosphorylase activities. AMRCs differ from PCs in: (1) a more intense reaction to the enzymatic activities ATPase, NADP, and succinic dehydrogenease, (2) a more intense immunostaining by AE1/AE3 and Ks4.62 anti-cytokeratin antibodies, and anti-estradiol receptor protein (D5) antibodies, and (3) a lower staining affinity for epithelial membrane antigen (EMA) antibodies. No positive immunostaining for the anti-cytokeratin Ks8.6 antibodies was observed in either AMRCs or PCs.     

The effect of immobilization on myotendinous junction: an ultrastructural,histochemical and immunohistochemical study

Kannus , P., Jozsa , L., Kvist , M., Lehto , M. & Järvinen , M. 1992. The effect of immobilization on myotendinous junction: an ultrastructural, histochemical and immunohistochemical study. Acta Physiol Scand 144 , 387–394. Received 28 April 1 991 , accepted 13 October 1991. ISSN 0001–6772. Tampere Research Station of Sports Medicine, UKK-Institute, and Department of Surgery, Tampere University Central Hospital, Tampere, Finland; Department of Morphology, National Institute of Traumatology, Budapest, Hungary; and Sports Medical Research Unit, Paavo Nurmi Center, University of Turku, Turku, Finland. The effect of immobilization on the myotendinous junction of the calf muscles in the rat was studied histochemically, immunohistochemically and morphometrically with a transmission electron microscope. After 3 weeks of immobilization, the contact area between the muscle cells and tendineal collagen fibres was reduced by almost 50% in both type I (slow-twitch) and type II (fast-twitch) muscle fibres. The terminal finger-like processes of the muscle cells became shallow and cylindrical or were completely atrophied. Their basal membranes were slightly thickened. Histochemically, the most remarkable alteration in the myotendinous junction was the marked decrease in the sulphate containing glyco-saminoglycans. In the basal lamina of the muscle fibres, the glycosaminoglycan and proteoglycan content was also reduced. Immunohistochemical analyses revealed that the amount of type III collagen was markedly increased on the myotendinous interface, but the amount and distribution of type I collagen was not affected by immobilization. These findings suggest that immobilization causes degenerative changes at the myotendinous junction, which, in turn, most likely decrease its tensile strength and may predispose it to rupture during activity.