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Biomedical subjects

L H Hoffman

Publications and source records attributed to L H Hoffman.

At least 55 records · Page 3Linked to original sources

Rapid method for preparation of nuclei from a mucin-producing solid tumor.

Nuclei were prepared from the human colon mucin-producing tumor GW-39. The method described involves incubation in beta-mercaptoethanol under mild conditions and results in the preparation of nuclei free of the mucin web that initially enmeshes them. A sensitive colorimetric immunoassay was used to confirm that nuclei prepared in this fashion were generally free of contaminating mucin. This method is mild, rapid, and should be applicable to other mucin-producing tumors.

Cell Fractionation↗

Attachment of pathogenic Neisseria to human mucosal surfaces: role in pathogenesis.

Studies of the interaction between Neisseria gonorrhoeae and human fallopian tube mucosa in organ culture suggest that attachment of gonococci is important, not only to secure th organism in the host, but also to initiate the disease process. The steps observed in gonococcal infection of fallopian tube organ cultures are: 1) attachment of gonococci to microvilli of nonciliated cells; 2) release from gonococci of lipopolysaccharide and possibly other toxic moities to cause mucosa damage; 3) engulfment or phagocytosis of gonococci by nonciliated cells; 4) transport of phagocytic vacuoles containing gonococci to the base of the nonciliated cells; and 5) exocytosis of gonococci within phagocytic vacuoles into the subepithelial tissues. In vivo, these steps might result in extensive local disease (e.g. salpingitis) or in the invasion of blood vessels to cause disseminated disease. Preliminary studies of human nasopharyngeal tissue in organ culture infected with Neisseria meningitidis indicate that meningococci attach to microvilli of nonciliated cells and are phagocytized by these cells. Meningococci subsequently appear in subepithelial tissues, though the route they take is not yet certain. These observations suggest at least some of the ways in which attachment may play a role in disease caused by N. gonorrhoeae and N. meningitidis. Mechanisms to block this attachment may provide new approaches to the prevention of infections caused by the pathogenic Neisseria.

Epithelium↗

Immunohistochemical localization of renin in mouse brain.

Immunohistochemical studies of mouse brain by the peroxidase-antiperoxidase method using monospecific antimouse renin antibodies has revealed the intracellular localization of renin in stellate and small ovoid cells. Renin-containing ovoid cells were observed in the spinal cord, medulla oblongata, pons, granular layers of the cerebellum, deep cerebellar region, and the lamina terminalis; whereas immunostainable stellate cells were found in the cerebral cortex, hippocampus, dentate gyrus and septum. Intracellular localization of renin rather than intravascular localization supports an endogenous origin of this enzyme in the brain. Wide distribution in different types of cells suggests different types of regulatory mechanisms. Double immunostaining with antigalactocerebroside and antirenin antibodies indicates the presence of renin in oligodendrocytes.

Animals↗

Effects of indomethacin on uterine capillary permeability and blastocyst development in rabbits.

Increased capillary permeability at implantation sites was demonstrated in rabbits by extravasation of intravascular blue dye on day 7 of pregnancy. Subcutaneous administration of indomethacin (Id, 8 mg/kg twice daily) on days 4-6 of pregnancy inhibited this uterine blueing response and appeared to reduce the size of implantation swellings. To test the latter observation blastocyst diameter and development of the embryonic disk were assessed at 144 hr post coitum. In females receiving indomethacin at the dose level which inhibited uterine blueing, blastocysts were significantly smaller than those from control females. Developmental staging of embryonic disks revealed only slight differences between the smaller (Id-treated) blastocysts and control blastocysts. No effect of Id was seen on ovarian function as judged by luteal weights and plasma progesterone and estradiol levels. Since the major biological effects of indomethacin are due to its inhibition of prostaglandin synthesis, it appears that prostaglandins may play a role in the uterine response to blastocyst stimulation and in the expansion of blastocysts in the rabbit.

Animals↗

Immunofluorescent localization of renin in mouse submaxillary gland and kidney.

An antibody prepared against purified submaxillary renin was used to determine the site of renin concentration in male mice using immunofluorescent localization. The results provide direct evidence that the granular tubules of the submaxillary glands are the source of submaxillary renin. The antibody against submaxillary renin cross-reacts with kidney renin as evidenced by immunofluorescent localization in the juxtaglomerular apparatus of mouse kidney.

Animals↗

Deciduogenic effect of prostaglandins in the pseudopregnant rabbit.

Intrauterine implants made of Silastic or wax and containing various additives, including prostaglandins (PGs), were used to characterize some of the factors involved in deciduogenesis in rabbits. Implants were inserted into uteri on Day 6 of pseudopregnancy and the response was assessed by light microscopy on Day 10. The surface area of decidual tissue in histological sections was employed as an index of the degree of decidualization obtained. PGE-2, PGF-2alpha and arachidonic acid each elicited a prominent decidual response, PGE-2 being most potent. Implants which released histamine or oestradiol-17beta had a very slight decidualizing effect in some females. Control implants and implants releasing dibutyryl cyclic-AMP had no deciduogenic influence. All implants, regardless of additive content, induced a symplasmic transformation of the uterine surface epithelium. It is concluded that PGs and the distension caused by the implant may be involved in deciduogenesis in the rabbit.

Animals↗

The effects of testosterone, 5alpha-dihydrotestosterone, 3alpha-androstanediol, and 3beta-androstanediol on epithelial fine structure of the rabbit epididymis in organ culture.

The fine structure of the corpus epididymidis of the rabbit has been studied following organ culture. Various modifications of tissue preparation and culture conditions were examined to obtain good maintenance of cellular integrity as well as to preserve sperm fertilizing ability. After 5 to 7 days in culture in the absence of hormonal support, the epididymal epithelium showed signs indicative of cellular regression. Such changes included shrinkage of the cells, loss of the border of stereocilia, decrease in smooth endoplasmic reticulum, and an increase in autophagic vacuoles. The presence of androgens in culture media prevented cellular regression to varying degrees, depending on the hormone utilized. With regard to maintenance of cellular integrity, potency of the androgens tested was as follows: 5alpha-dihydrotestosterone greater than or equal to 3alpha-androstanediol greater than testosterone greater than 3beta-androstanediol. Addition of insulin to dihydrotestosterone-containing cultures resulted in no improvement in maintenance. Phagocytosis of spermatozoa by epithelial cells was observed in cultured tubules and the degree of spermiophagy was inversely proportional to successful maintenance of fine structural characteristics of epithelial cells. The morphological findings reported here correlate well with the fertilizing ability of spermatozoa from cultured epididymis as reported in an accompanying communication.

Androstane-3,17-diol↗

The effects of testosterone, 5alpha-dihydrotestosterone, 3alpha-androstanediol, and 3beta-androstanediol on the maturation of rabbit epididymal spermatozoa in organ culture.

The fertilizing ability of spermatozoa from epididymal tubules maintained in organ cultures from 1 to 7 days was assessed after artificial insemination into receptive does. It was found that spermatozoa from the distal corpus which were already capable of fertilizing eggs prior to the cultures retain this ability for 1 day without addition of hormone and for 3-4 days when testosterone (0.5 mug/ml) or 5alpha-dihydrotestosterone (0.5 mug/ml) is added to the culture medium. Spermatozoa from the proximal corpus which were not capable of fertilizing eggs prior to the cultures remain so after 1 day in cultures without addition of hormone. Testosterone, 5alpha-dihydrotesterone, 3alpha-androstanediol, or 3beta-androstanediol was added to cultures of proximal corpus at a concentration of 0.5 mug/ml. Only with 5alpha-DHT is the mean percentage of fertilization significantly higher than the percentage obtained without addition of hormone. Insulin does not potentiate the effect of 5alpha-DHT on sperm fertilizing ability. Epithelial growth factor is ineffective. Spermatozoa from the caput epididymidis kept in cultures for 1 to 4 days remain infertile. The results are discussed in light of the morphological findings presented in the preceding communication and in relation to the physiological requirement for sperm maturation in the epididymis.

Androstane-3,17-diol↗

Studies on the progestational endometrium of the rabbit. II. Electron microscopy, day 0 to day 13 of gonadotrophin-induced pseudopregnancy.

The fine structure of the endometrial epithelium of the pseudo-pregnant rabbit from the day of induced ovulation (day 0) to the 13th day is here correlated with previously defined light microscopic phases. In Phase 1 (0-1 day), in which there is a presumed "priming" of the endometrium by ovarian steroidal hormones, no changes were observed. In Phase 2 (1-3 days), in addition to mitotic activity, the epithelium showed a disappearance of the mucification and lymphocytic migration typical of Phase 1 and also of the non-pregnant or "estrous" phase, and showed other nuclear and cytoplasmic changes which probably reflect endogenous growth and protein synthesis. In Phase 3 (4-6 days), two distinct populations of reacting cells were present: (1) surface and cryptal cells investing the now folded mucosal surface, and (2) glandular cells. The first group showed characteristic dome-like protrusions of the cytoplasm into the lumen, and also showed distinct cytoplasmic and nuclear changes which appear to be a prelude to the succeeding phase of fusion but are not necessarily secretory in type. The glandular cells, in contrast, showed cytoplasmic changes which appear to reflect active secretory activity (hypertrophy of the Golgi area, cytoplasmic vacuoles containing electron-opaque material, etc.). This phase coincides with the maximal secretion of uterine-specific proteins, and electron-opaque material is abundant within the endometrial lumen. In Phase 4 (6-8 days), the surface and cryptal epithelium undergoes a transformation into multinucleated cells, the result of a process of lysis of intervening plasma membranes, the precise mechanism of which (i.e., with or without initial membrane fusion) was not determined. Cell fusion proceeded earlier and more actively mesometrially than antimesometrially. The glandular cells showed evidence of reduced secretory activity, but did not at any stage undergo multinucleate-cell transformation. In Phase 5 (8-13 days) there was progressive fusion, and the number of nuclei per cytoplasmic sac appeared increased, presumably due to the continued action of progesterone which is maximal during this phase. Glandular cells showed further reduced secretory activity but remained columnar. Ciliation of the epithelium was sporadic in the pre-secretory phases and rare or absent in the secretory and fusion phases; it became widespread during the phase of decline after day 14, a period which will not be included in this study. The fine structure of the ciliated cells was the same at all stages; there was no evidence for their origin from a reserve population; it is possible that they arise by modification of the multinucleated cells. Cytoplasmic crystals and intramitochondrial densities or lamellae were observed during the secretory and fusion stages, the former only in the glands, the latter in the surface and cryptal epithelium. They appear to be associated with rising or maximal progesterone secretion.

Animals↗