Walter H. Wilborn

Scanning electron microscopy of human spermatozoa after incubation with the spermicide nonoxynol-9

The most frequently utilized spermicide in vaginal contraceptives is No-9. Schill and Wolff used TEM to demonstrate the focal effects of No-9 on human sperm and reported that No-9 damaged cell membranes and acrosomal membrane complexes. The present study by SEM was made to assess the extent of membrane damage due to the direct action of No-9 during an incubation period of only 5 minutes. SEM revealed that No-9 caused loosening and detachment of acrosomal, neck, and midpiece membranes of all sperm even at the lowest concentration tested (0.05%). The severity of membrane alterations observed in any of these regions would render sperm immotile and unable to penetrate the ovum. The fact that these alterations are produced within 5 minutes after exposure to No-9 attests to the effectiveness of No-9 as a vaginal contraceptive.

Histologic classification and pathologic findings for endometrial biopsy specimens obtained from 2964 perimenopausal and postmenopausal women undergoing screening for continuous hormones as replacement therapy (CHART 2 Study)

OBJECTIVE The aim of this study was to screen for disorders and to histologically classify endometrial biopsy specimens from 2964 perimenopausal and postmenopausal women who were candidates for hormonal replacement therapy. STUDY DESIGN Endometrial biopsy specimens were obtained with a Vabra aspiration curette, processed by standard methods, and stained by hematoxylin and eosin and special methods to reveal subtle features of the endometrium. RESULTS Of the endometrial biopsy specimens, 68.7% were atrophic, 23.5% were proliferative, 0.5% were secretory, 0.6% were hyperplastic, 0.07% were adenocarcinoma, and 6.6% were insufficient for classification. Three independent senior microscopists agreed on the classification of each biopsy specimen. CONCLUSION The number of patients is the largest ever screened for a single hormone replacement therapy study. The low yield of endometrial cancer indicates that biopsies are unnecessary before hormone replacement therapy is initiated in asymptomatic women.

Tetrazolium artifactually indicates superoxide dismutase-induced salvage in reperfused rabbit heart

We tested the ability of a single dose of superoxide dismutase to induce salvage of reperfused rabbit myocardium. Infarct size was measured by tetrazolium method following 3, 24, or 72 h of reperfusion. In addition, the 24 h reperfused hearts were examined to determine if the drug induced salvage in those hearts was reflected in the histology. A coronary arterial branch was occluded for 45 min and then allowed to reperfuse for 3, 24 or 72 h. At the end of the reperfusion period the hearts were removed, perfusion stained with triphenyl tetrazolium, and fixed in buffered formalin. The hearts were sectioned and infarct size was determined in all groups. In addition, the 24 h heart slices were prepared for histology with H&E staining. The results revealed that 5 mg/kg hSOD treatment was associated with smaller infarcts in the 3 and 24 h groups but that differences were no longer apparent in the 72 h group. The 24 h control hearts showed good correlation between infarct size by TTC and that by conventional histology. In the 24 h treatment hearts, however, infarcts by TTC averaged only about 1/2 the size of those by conventional histology. We conclude that a single dose of hSOD fails to offer a sustained reduction of infarct size. Furthermore, histology from the 24 h reperfused group revealed that hSOD did not delay the onset of necrosis but rather simply caused dead tissue to retain its ability to reduce the tetrazolium salts.

Superoxide dismutase conjugated to polyethylene glycol fails to limit myocardial infarct size after 30 min ischemia followed by 72h of reperfusion in the rabbit

We tested whether recombinant human superoxide dismutase conjugated to polyethylene glycol (PEG-SOD) to prolong its plasma retention time could limit myocardial infarct size in an ischemia-reperfusion model in the rabbit. One group of animals received 1000 units/kg of PEG-SOD as an intravenous bolus 15 min before coronary occlusion. A second group received saline only and served as controls. Under pentobarbital anesthesia, a left coronary branch was occluded for 30 min and then reperfused. The surgical wounds were repaired and the animals were allowed to recover. Seventy-two hours after the coronary occlusion, the heart was excised and the size of the area at risk (ischemic vascular bed) was assessed with fluorescent particles and the infarct size was determined by histology (Hematoxylin-eosin, Azan stain). Infarct size as a percentage of the area at risk was similar between the groups, 46.5 + 2.7 in the PEG-SOD group (n = 8) and 48.9 + 3.1 in the control group (n = 8). There were no significant differences between the groups indicating that PEG-SOD did not limit infarct size in this model.

Ultrastructural Identification of Langerhans Cells in Seborrheic Keratoses

Lesions of seborrheic keratoses studied by transmission electron microscopy showed an unusually large number of Langerhans cells in the epidermis. The role of Langerhans cells in the development of these lesions needs further study.

Ultrastructural Basis for the Assay of Topical Acne Treatments. Transmission and Scanning Electron Microscopy of Untreated Comedones

After two decades of using systemic antibiotics as a main treatment for acne, emphasis is again being placed on topical agents. Thus, it is highly desirable to have a procedure whereby the activity of the various compounds can be evaluated by direct visualization. Scanning electron microscopy combined with transmission electron microscopy provide the tools for such an assay. This study describes the ultrastructure of untreated comedones and provides the baseline or control data necessary for testing topical treatments. Comedones obtained by punch biopsies or comedo extractors were processed for electron microscopy and studied with an ETEC Auto‐scan Scanning Electron Microscope and with Philips EM 300 and EM 301 Transmission Electron Microscopes. Microorganisms, keratinized cells, sebum, and hairs interact with each other to form the comedonal mass. Corynebacterium acnes and Pityrosporum ovale proliferate abundantly in close association with sebum and penetrate the keratinized cells. Fine structural details of bacteria and yeasts as well as features of host‐microbial relationship have been elucidated.

A large parosteal osteosarcoma with transformation to high‐grade osteosarcoma. A case report

This case of a large (31 cm) parosteal osteosarcoma had x‐ray documentation 4 1/2 years before treatment. When the tumor began enlarging rapidly, treatment was sought. The tumor histologically showed typical parosteal osteosarcoma as well as malignant cartilage, osteoid, and undifferentiated sarcoma. The case shows one course a parosteal osteosarcoma can take over a 7‐year untreated period. The significance of high‐grade sarcoma and intramedullary involvement is discussed. When unequivocal high‐grade osteosarcoma is present, it should be treated with radical resection and adjuvant multi‐drug chemotherapy.

Antiproliferative effects of low-dose micronized progesterone*

OBJECTIVE To study the pharmacodynamic effects of oral micronized P on endometrial maturation. DESIGN This was a controlled, open, parallel group, pilot study. SETTING The experiment was performed in an outpatient academic clinical research unit. PATIENTS Twelve healthy, P-challenged, estrogen-primed, postmenopausal women participated in the study. INTERVENTIONS Patients were given 300 mg micronized P daily (8:00 A.M.) or twice (8:00 A.M. and 4:00 P.M.) daily from study days 1 through 14 after estrogen priming for 30 days. Blood samples were taken at 0, 0.5, 1, 1.5, 2, 3, 4, 6, and 8 hours after the 8:00 A.M. dose on study day 1 and 14 and again at 8:00 and 9:30 A.M. on days 3 and 5 fasting, days 7 and 9 after a fatty meal, and day 11 after a high fiber meal. Endometrial biopsies were taken on day 1 and 14. MAIN OUTCOME MEASURES Progesterone concentrations were measured. Endometrial biopsies were studied for effects on histology, glycogen content of glands, ribosomal RNA, and nuclear estrogen receptors in glands, surface epithelium, and stroma. RESULTS Day 1 and 14 P kinetics were similar for 8 hours. Dose-dependent increases in glandular glycogen, decrease in ribosomal RNA, and decrease in nuclear estrogen receptors were demonstrated. CONCLUSIONS Oral micronized P can induce antiproliferative changes in the human endometrium at doses lower than those required for transformation of the endometrium to a full secretory state.

Low strength anthralin in psoriasis.

Anthralin in low concentrations (0.01%–0.03%) was applied in an ointment form once per day in the hospital to 130 psoriasis patients. The treatment program included also the use of UVB light, emollients and bath oil and an antihistamine. Clearing of psoriasis was achieved in all patients in an average time of 4 weeks. The involution of the lesions was carefully studied by correlated clinical observations, light microscopy, transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Striking healing of the holes in the stratum corneum and of the gaps in the basement membrane occurred. Low strength anthralin, in addition to its effectiveness, did not produce irritation, staining or side‐effects.

Association of vitiligo with Sjögren's syndrome.

Sir, Many years after publication of the last comprehensive treatise on vitiligo (1), two recent books (2, 3) provide a thorough review of the literature on this common autoimmune disorder. Interestingly, while vitiligo may be associated with one or several of many systemic abnormalities (2), to our knowledge there have been no reports of its association with Sjögren’s syndrome, another autoimmune condition. In our Vitiligo Unit we observed two women (32 and 35 years old) first affected by Sjögren’s syndrome followed 2 – 3 years later by generalized vitiligo. Onset of the two conditions took place after implantation of a copper intrauterine device (IUD), a frequent, although still unexplained finding among our vitiligo patients. Lymphocytic infiltration and destruction of the lacrimal gland epithelium is a main histologic feature of Sjögren’s syndrome (4). Lymphocytic infiltration of the dermis and destruction of melanocytes is a hallmark of vitiligo. In Sjögren’s syndrome, 30% of patients have moderate leucopenia (5). The leucocyte counts in our two patients were also diminished (3400 and 5400/mm). Both disorders are associated with an increased incidence of autoantibodies (2, 6). Among the various pathogenetic mechanisms for these two entities, nutritional deficiencies have been shown to play a role. Vitiligo may be accompanied by deficiency of vitamin B12, folic acid, vitamin C, vitamin B6, vitamin E, iron or copper (2). Sjögren’s syndrome can develop as a unique characteristic of scurvy (7), or as a manifestation of vitamin A deficiency (8). In patient 1, there were diminished serum levels of folic acid and vitamin B6. In patient 2, the serum levels of folic acid and vitamin B6 were near the low limit of the normal range (3.9 and 6 ng/ml, respectively). Immunological abnormalities may result from nutritional deficiencies in general or from deficiencies of single elements such as minerals and vitamins (9); they might thus have played a role in the autoimmune disorders of these two patients. After the most commonly associated abnormalities have been ruled out when evaluating vitiligo patients, a search to determine the presence of Sjögren’s syndrome seems warranted. Likewise, the skin of patients with Sjögren’s syndrome should be carefully examined both by direct clinical inspection and under Wood’s light to detect areas of depigmentation, which are always discernable in vitiligo.