Isolation of highly purified hepatocytes from adult pig liver in multivisceral organ procurement.
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Biomedical subjects
Publications and source records attributed to J Codesal.
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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.
125I-glucagon binding sites have been detected and quantified on the intestinal smooth muscle cells of the dog by means of ultrastructural autoradiographic methods. These binding sites are located mainly in the plasmalema. The present findings established a morphological correlation with the physiological data concerning the spasmolytic function of the glucagon on the intestinal wall.
The testes of stillborn fetuses (from 13 to 28 weeks of gestational age), fetuses born alive (from 29 weeks of gestational age) who died a few days later, and infants dying 1 to 8 months after birth were processed for light and electron microscopy. Paraffin-embedded material was stained with the avidin-biotin peroxidase complex (ABC) method for immunohistochemical detection of testosterone (T) in order to quantify the age-related changes in the number of T-positive interstitial cells. This number decreased progressively from the 24th week of gestation up to birth and remained unchanged up to the second month of postnatal life. During the third month of age, the number of T-positive cells rose markedly but fell again from the fourth month to the end of the study. The ultrastructural study revealed the following types of interstitial cells at all ages studied: fibroblast-like cells, myofibroblast-like cells, developed fetal Leydig cells, degenerating fetal Leydig cells and infantile Leydig cells with a multilobed nucleus and focal cytoplasmic accumulations of smooth endoplasmic reticulum and lipid droplets. Quantitative ultrastructural studies revealed that the changes in the number of fetal Leydig cells with age were similar to those found in the number of T-positive cells although, for each age studied, absolute values were higher in the ultrastructural study. The number of infantile Leydig cells increased with age.
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Of 723 infertile men (128 with a history of cryptorchidism) whose testes were biopsied at the outer lateral face of the testis, five presented carcinoma in situ (CIS) in one testis. These testes were removed, serially sectioned, and examined by light microscopy. In order to evaluate whether only one or two biopsies are sufficient to diagnose CIS, before sectioning the testes four biopsies were taken at the anterior face, posterior face, superior pole, and inferior pole of the testis, respectively. Two of the five men had undergone orchiopexy in infancy and the testis contained tubules with Sertoli cells and isolated spermatogonia. CIS was also present in some tubules that were principally located near the rete testis. Of the four simulated biopsies, only that performed at the posterior face of the testis revealed CIS. The other three infertile men showed tubules with complete, although reduced, spermatogenesis, and tubules lined by Sertoli cells only. CIS was found in both types of tubules. These tubules with CIS formed lobules that extended throughout the testicular parenchyma. Most simulated biopsies performed in these three testes showed CIS. The average nuclear DNA content of CIS cells was about 4c in all testes. This content was similar both in tubules with complete spermatogenesis and in tubules with Sertoli cells only.
The proliferative activity of human spermatogonia from the fetal period to senility was studied by means of cytophotometric quantification of the nuclear DNA content in histological sections. Proliferating spermatogonia that were replicating or had replicated their DNA (DNA content between 2.5c and 4.5c) were observed in all ages. The percentages of these spermatogonia were high (18.2%) in the second trimester of gestation, decreased in the third trimester (8.2%), maintained similar values in newborns (7.1%) and infants (9.5%), and increased markedly in 4- to 9-year-old children (22.5%). The latter percentage was maintained during puberty (20.1%), decreased significantly in adulthood (17.8%), and was higher in aging testes (25.2%). About 2% of spermatogonia with a DNA content higher than 4.5 c were observed from 4-15 years of age as well as in the testes of elderly men. Sertoli cells replicating their DNA were observed only in fetuses.
A histochemical and ultrastructural study of the testes, epididymides and spermatic cords from 6 adult men with amyloidosis secondary to several diseases is described. Amyloid fibrils, consisting mainly of AA protein, were deposited in the walls of blood vessels of the testis, epididymis and spermatic cord, as well as in the tunica propria of the seminiferous tubules. The seminiferous epithelium showed slight hypospermatogenesis. Many Leydig cells exhibited unstained 'bar' or 'arc'-shaped cytoplasmic areas similar to the cholesterol deposits of Leydig cells in adrenoleukodystrophy.
Hepatocyte transplantation (HT) is among the most frequently employed approaches in the treatment of liver disease. This work deals with the occurrence of rejection when allogeneic hepatocytes are transplanted into the spleens of rats. Two strains of rats were used: Lewis and F344. The recipients were divided into two groups, one of which received ciclosporin (Cic), while the other received no treatment. After HT, the spleens were removed periodically. The animals that did not receive Cic suffered such intense rejection that 7 days after HT no hepatocytes could be identified in the splenic pulp. On the other hand, hepatocytes could be observed throughout the entire study in the spleens of rats treated with Cic. We conclude that HT is followed by intense rejection, which can be avoided by the administration of Cic.
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The numbers of each different cell type in the human seminiferous epithelium were determined throughout the 6 stages of the cycle in both semithin and ultrathin sections obtained from 15 young adult men with normal testicular histology. Up to 4 types of A spermatogonia (Ad, Ap, Al and Ac) were distinguished. In addition, the DNA nuclear content of seminiferous epithelium cells was determined on Feulgen-stained sections. Both Ad and Ap spermatogonia showed a 2c DNA content and were present in the 6 stages of the cycle, though their numbers decreased in stages III-V. Both Al and Ac spermatogonia showed a DNA content varying from 2c to 4c. Al spermatogonia were observed in stages III-V; their numbers plus those of Ad spermatogonia in these stages were similar to the numbers of Ad spermatogonia in the other stages lacking in Al spermatogonia. Ac spermatogonia appeared in stages III-VI and their numbers plus those of Ap spermatogonia in stages III-V were similar to the numbers of Ap spermatogonia in the other stages lacking in Ac spermatogonia. The results suggest that Ad spermatogonia are the stem cells. Some of them replicate their DNA; during this replication they appeared as Al spermatogonia. Al spermatogonia divide, giving rise to both Ad and Ap spermatogonia. Some Ap spermatogonia replicate their DNA; during this process they are transformed into Ac spermatogonia which divide, giving rise to B spermatogonia.
A new method for correlating testicular biopsies and spermiograms is proposed. The number of spermatogonia, round spermatids, and elongated spermatids per cross-sectioned tubule were calculated in the testes from 33 subfertile men and in 10 control normal testes. According to these quantitations, the testes in subfertile men were classified as testes with maturation arrest of spermatogenesis, testes with hypospermatogenesis, and testes with associated maturation arrest and hypospermatogenesis. A power regression curve correlating the number of elongated spermatids and sperm numbers in the spermiogram was performed.
A comparative morphologic study between the testes of 25 young adult men and 41 elderly men without testicular or related pathological conditions revealed the presence of multinucleate spermatids, showing up to 86 nuclei, in the testes of 3 of the elderly men. The formation of multinucleate spermatids is probably due to cell fusion, since the number of nuclei in these cells is not always 2n and multinucleate spermatocytes were uncommon. This anomaly seems to be another manifestation of an involutive process that also affects other cell types in the aging testis.
The numbers of Ap and Ad spermatogonia per unit section of the testis were calculated in autopsy specimens from young adults and elderly men without testicular pathology. The number of Ap spermatogonia decreased from the 6th decade of life, whereas that of Ad spermatogonia began to decrease in the 8th decade. Although it has been reported that Ad spermatogonia are more sensitive to noxious agents than Ap spermatogonia, the involution of Ap spermatogonia precedes that of Ad spermatogonia. These findings provide new information on concepts relating to the spermatogonia precedes that of Ad spermatogonia. These findings provide new information on concepts relating to the spermatogonial stem cell in man.
A solution of lead acetate (300 mg/L) was administered via drinking water to pregnant Wistar rats from day 1 of pregnancy to delivery (Pb-treated day 0 group) or throughout gestation and early lactation (from day 1 to day 5 postnatal) (Pb-treated day 5 group). When the pups were born, four dams and their offspring in each group (control day 0, Pb-treated day 0, control day 5, and Pb-treated day 5) were sacrificed on day 0 (day 0 groups) or on day 5 (day 5 groups). Relative testicular weight and gross testicular structure were not altered by the treatment. The seminiferous tubule diameter and the number of prospermatogonia were reduced by the treatment. Determination of the n-ploidy stage of prospermatogonia indicates that these cells have more proliferative activity in Pb-treated rats than in control rats. On the other hand, the total DNA, RNA, and protein content of the testes in treated rats was significantly reduced, but the DNA: RNA ratio remained unaltered.
The number of mononucleate and multinucleate Leydig cells per unit area of the testis was determined in normal adult men using the peroxidase-anti-peroxidase method for testosterone detection. The results of this study indicate that the number of multinucleate Leydig cells increases markedly with age, whereas the total Leydig cell population decreases.
A cytophotometric quantification of DNA in tumor cells was performed in histological sections of orchidectomy specimens from 36 men with testicular germ cell tumors (TGCT), 7 of them showing more than one tumor type. Among the variants of seminoma (classic and spermatocytic) the lowest DNA content were in spermatocytic seminoma. With respect to non-seminomatous tumors (yolk sac tumor, embryonal carcinoma, teratoma, and choriocarcinoma), choriocarcinomas showed the highest DNA content, and the lowest value was found in teratomas. No significant differences were found between the average DNA content of seminomas (all types) and non-seminomatous tumors (all types). Both embryonal carcinoma and yolk sac tumor showed similar DNA content when they were the sole tumor and when they were found associated with other tumors. In this study, except for the 4 cases of teratoma and the case of spermatocytic seminoma, all TGCT examined did not show modal values of DNA content in the diploid range. Such an elevated frequency of aneuploidism in these tumors may be helpful for their diagnosis.