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Androgenetic Alopecia (AA) afflicts a large part of the population and of the many treatments available today none is completely satisfactory. Testing the efficacy and safety of a novel topical treatment for AA which is based on cell culture medium supplemented with insulin, thyroxin and growth hormone (CCM). The 48 participants classified as androgenetic alopecia Type II, III or IV on the Hamilton scale, concluded a randomized, vehicle-controlled, double-blind trial of 6 months duration. Under occlusive cover the gel was self applied for at least 3 hours daily. Evaluation was based on hair counts, investigator global assessment and participants self-administered questionnaire. Cessation of hair loss was reported by most participants within 28 weeks, and further confirmed by the hair count (HC) in ~80% of participants. Moreover, as early as 4 months after the start of the treatment, a time dependent increase of up to 50% in HC was observed. The average change in HC between the two groups differed significantly (p=0.007), with values of 4.1% for control and 13.8% for CCM. Following 4 months of treatment, a time dependent increase in HC (>10%) above minimal was observed in 55% of the CCM and 25% of the control and this trend continued. At 6 months 63% of the CCM and 33% of the control group exhibited increase of HC higher than 10%. The average increase in HC in the CCM and the control groups was 17.1% and 8.9% respectively (p=0.035). Self evaluation questionnaires revealed a time dependent increase in satisfaction in the CCMusers compared to the control. While the average score at T2 was similar in CCM and control (2.7 and 2.6 respectively), the score at T6 in the CCM increased to 5.9 and decreased to -0.4 in the control (p=0.007). Global-clinical evaluation following six months treatment revealed significantly (p=0.02) more hair loss in the control group (40%) compared to the CCM (7%) treated group. CCM was found effective in treating androgenetic alopecia in men. It induced cessation of hair loss, increased rate of hair growth and appearance of new hair. No side effects were reported or observed.

This study was designed to test the efficacy of enriched cell culture medium as a wound dressing. The rationale was to create within the wound space an optimal microenvironment, conducive to cellular proliferation, vascular granulation tissue formation, and epithelialization. This study was performed on various wounds that failed to respond to previous conventional treatments.A total of 288 wounds were within the inclusion criteria, with only contaminated and neoplastic wounds excluded. Most of the patients (80 percent) were ambulatory, and the wounds were examined by the attending physician once every 7 to 14 days at an outpatient clinic. The remaining 20 percent of patients were admitted to the study while hospitalized. Cell culture medium MCDB, supplemented with insulin, thyroxin, and growth hormone, was gelled. The gel was self-applied once a day to freshly washed wounds, covered with a gauze pad, and anchored with netting. Healing started 7 to 14 days after the initiation of treatment with enriched cell culture medium. However, the criterion for success of the treatment was determined on complete wound closure, which was achieved in 189 of 288 wounds (65.6 percent). Wound closure was correlated with the initial wound volume, stage, and origin. The average time required for closure of wounds caused by systemic pathologies (n = 181) and those based on regional status (n = 107) were 12.0 and 4.4 weeks, respectively, compared with 290 and 10.3 weeks of the previous conventional treatment. In 19 extensive wounds, when vascularized granulation tissue was established, a successful surgical closure was attained. Most wounds of patients who did not continue the enriched cell culture medium treatment (34.4 percent) manifested reduced wound volume, ranging from 11 to 98 percent of initial volume. Discontinuation of treatment was associated with difficulties in reaching the clinic for the weekly examination, rather than for reasons directly related to the treatment itself, and occurred significantly earlier during the treatment period.Thus, enriched cell culture medium was effective in stimulating wound healing in recalcitrant wounds. The healing was rapid with minimum scarring and pain. No side effects or allergic reactions were reported or observed.

Problem: The purpose of this study was the isolation and characterization of decidual extract proteins that exhibit mitogenicity on endothelial cells., Method of Study: A partially purified extract (F1 fraction) was obtained from human decidua of the first trimester of pregnancy. F1 was separated by heparin-sepharose column and showed significant mitogenicity on bovine brain capillary endothelial (BBCE) cells in vitro, using methylene blue stain nuclear assay. Sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed four fractions with MW less than 30 KDa., Results: Mitogenic fraction, E11-12 (eluted at 1.5 M NaCl) was identified as basic fibroblast growth factor (bFGF). Vascular endothelial growth factor (VEGF) and acidic (a)FGF were not identified as one of the mitogenic fractions. However, fractions E5-6, and E7-8 induced statistically significant mitogenicity at concentration of 1 pg/mL, which is 10(3) times lower than bFGF. Sequencing of E5-6 fraction identified NKG5 protein, a putative secreted polypeptide derived from human natural killer (NK) cells and activated T cells of unknown function., Conclusion: This work demonstrates that NKG5 stimulates mitogenicity of endothelial cells and may be involved in angiogenesis.

Autologous fat grafting as a technique to correct soft-tissue defects is a controversial subject. The high percentage of fat resorption and the resulting need for additional grafting considerably reduce the value of this method. The purpose of this study was to evaluate the clinical application of tissue-culturing methodology in the handling of the lipocyte aspirate in an endeavor to improve the survival rate and therefore the take of the grafted lipocytes. The method consists of syringe aspiration of the lipocytes from the donor site, isolation of intact lipocytes by gentle centrifugation, suspension of the aspirate in an enriched cell culture medium, and injection of the cell suspension into preformed subdermal tunnels. A number of media were tested and shown to prolong the survival of lipocytes ex vivo using fluorescent acridine orange stain. Implementing the integrated cell culture techniques increased the lipocytes' viability, as indicated in clinical evaluation in which the amount of graft take ranged between 50 and 90 percent. The results of 15 patients with varied types of cases who were operated on using this new methodology show that the tissue defect was filled and remained so in postoperative follow-ups of 6 to 24 months. A three-dimensional CAT scan-aided evaluation method was developed and used in one of four case histories presented herein.

Recently, the superficial musculoaponeurotic system (SMAS) was found to be a composite tissue comprising collagen, elastic fibers, and fat cells in an extracellular viscous matrix. Both SMAS and facial skin tissues exhibit viscoelastic properties, but SMAS tissue has delayed stress relaxation. As a consequence, SMAS is viewed as a firmer elastic foundation for the more viscous facial skin. In some patients, a slackening effect of SMAS tissue takes place over a period ranging from weeks to months after tightening. To determine the relative quantity of viscoelastic components and better understand their biomechanical behavior, a quantitative morphometric study of the elastic and collagen fibers in the SMAS and facial skin was conducted. Thirty-four SMAS preparations were taken from 17 patients during either primary face lift operations (12 women) or reoperative face lift procedures (4 women, 1 man), which were performed 4 to 9 months after the original surgery, to examine the elastin and collagen content. For comparison, preauricular skin was also gathered from these patients. The specimens were stained with Weigert's staining to identify elastin and collagen fibers. Using a computerized morphometric analysis, 100 fields of each SMAS and skin specimen were examined. According to our findings, the average percentage of elastin and collagen fibers in SMAS and facial skin was as follows: (1) the percentage of elastin fibers in the SMAS was 4.71 +/- 1.2 (standard error of mean +/- 0.0291); (2) the percentage of elastin fibers in the skin was 6.1 +/- 1.8 (standard error of mean +/- 0.0436); (3) The percentage of collagen fibers in the SMAS was 38.7 +/- 5.9 (standard error of mean +/- 0.1430); and (4) the percentage of collagen fibers in the skin was 48.47 +/- 6.96 (standard error of mean +/- 0.1688). A statistical significance of p < 0.0001 was demonstrated between the collagen and elastin groups. A different percentage of elastin and collagen fibers was found among the 17 patients and within each of them separately. Neither gender nor age differences were found regarding elastin and collagen fiber content. No statistical differences were demonstrated between specimen sources, i.e., whether the operations were primary or reoperative face lift procedures. Findings from previous studies indicate that the cheek has two viscoelastic layers, the skin and the SMAS. The proportional similarity in average percentages of elastin and collagen in SMAS and facial skin cannot explain the relatively delayed stress relaxation effect of the SMAS. Therefore, the fat cells that are found exclusively in the SMAS probably lend a certain degree of firmness to this layer and play a significant role in the long-term efficacy of SMAS surgery.

Injection of aspirated fat is now the most commonly used technique for the filling of depressed areas. Partial absorption of the injected fat is the main limitation of this procedure. Cariel T.M. is an enriched serum-free cell culture medium, its ability to enhance the survival of human aspirated fat grafts was investigated in the nude mouse model. A volume of 0.75-cc Cariel preprocessed fat was injected under the scalp skin of 16 nude mice in the experimental group, and the same volume of saline preprocessed fat was injected to 15 control group of mice. Significant maintenance of the weight, 46 percent in the experimental group compared with 29 percent in the control group (p < 0.008), and the volume, 44 percent in the experimental group compared with 31 percent in the control group (p < 0.026), was observed, after 15 weeks, in this newly used model. It seems that addition of the nutrients enriched with anabolic hormones enabled the survival and take of more adipose cells in the graft.

A study was undertaken to determine the physical properties and microscopic structure of the superficial musculoaponeurotic system (SMAS) tissue. Forty virginal specimens and eight reoperated specimens were examined. The following findings were discovered. 1) Microscopic appearance shows the SMAS to consist of collagen fibers, a relatively high concentration of elastic fibers interspersed with fat cells. 2) On scanning electron microscopy, the virginal SMAS shows the collagen fibers to have a similar convoluted appearance as in the dermis. There is some evidence of parallelization of the collagen fibers in the reexcised SMAS specimens. 3) Mechanical testing (Instron) demonstrates that both the SMAS and preauricular skin were subjected to a series of loading/ unloading tests at various rates, amplitudes, and stress relaxation tests. Both sets of specimens indicated definite viscoelastic properties. Although the mechanical behavior of both tissues was somewhat similar, the viscoelastic effect of the SMAS was less pronounced. A slackening effect of the SMAS indicated a gradual expansion of the SMAS postoperatively. These results could provide some indication of the long-term effects of SMAS surgery.

Because of the widespread reliance on SMAS tightening procedures in present-day face lift surgery, a study was undertaken to examine the physical properties and microscopic structure of both virginal (40 specimens) and reoperated (8 specimens) SMAS tissue. The findings could be of practical value to the surgeon and are reported herewith: First, the SMAS is a composite fibrofatty layer comprising collagen and elastic fibers interspersed with fat cells. Second, microscopic appearance shows a considerable amount of elastic fibers in close relationship to the collagen fibers. Third, on scanning electron microscopy, the collagen fibers in the virginal SMAS show a convoluted appearance similar to that found in the dermis. In the reexcised SMAS tissue, there is some evidence of parallelization of the collagen fibers as seen in the stretched dermis. Fourth, mechanical testing (Instron), i.e., a series of loading/unloading tests at various rates and amplitudes, and stress relaxation tests were performed on samples of preauricular skin and SMAS. These indicated definite viscoelastic properties for both sets of specimens, with the tendency of an increased stiffness and a reduction in viscoelastic effects on repeated working of the samples. Overall, the mechanical behavior of both tissues was somewhat similar, the viscoelastic effects in SMAS being less pronounced. A nonlinear viscoelastic model is under development to represent the behavior of both tissues. The implications of these results may help to explain the slackening effect observed in some postoperative patients.

Among the current methods employed in the treatment of wounds, a moist dressing is considered to be the optimal environment for the process of healing thereby avoiding dessication of the wound bed. This study is based on the hypothesis that wound cell proliferation is dependent not only on moisture but also upon the composition of the moist microenvironment in the wound. That composition in turn is formed by diffusion of nutrients from the existing vascular bed in and around the wound as well as by the wound cells' cellular products. Since in wounds the impaired vascular supply causes tissue deprivation, a continuous supply of nutrients and hormones will create an optimal substrate for cellular mitogenic activity, synthesis of matrix, growth factors and cytokines leading to wound healing. Modified serum-free cell culture medium was supplemented with non-steroidal anabolic hormones: growth hormone, thyroxin and insulin, transferrin and sodium selenite. The medium was prepared in a 1 per cent alginate gel matrix. Under general anaesthesia with ketamine (Imalgene 1000, Rhone Merieux, France) four 2 x 2 cm full-thickness skin patches were surgically extirpated from the dorsum of Hartley-derived guinea-pigs. Each experimental group consisted of seven animals, i.e. 28 wounds that received the same treatment. Compositions of gelatin in saline, agarose in saline, agarose in medium and agarose in saline supplemented with the three hormones were compared to agarose in medium supplemented with the three hormones. After application of the gel (1 ml/cm2), the wounds were dressed with gauze, elastic adhesive bandage and netting.(ABSTRACT TRUNCATED AT 250 WORDS)

Human decidual capillary endothelial (HDCE) cells, obtained from decidual fragments of legally induced first-trimester terminations of pregnancies, were cultured in a serum-free medium supplemented with human uterine angiogenic factor (HUAF). The method of isolating the cells from the decidual tissue is described. Identification of the decidual endothelial cells was based on light- and electron-microscopic observations as well as on antifactor VIII immunoperoxidase-staining technique. The HDCE cell culture in the serum-free medium lasted for 25 weeks through eight subcultures. The cells frozen in glycerin yielded 80% viability after thawing. Electron-microscopic observations of the monolayer demonstrated Weibel-Palade bodies, desmosomes and tight junctions. HUAF is mitogenic to HDCE at 10-100 ng/ml. The dependency of HDCE cells grown in culture on HUAF may explain in part the mechanism involved in the dynamic vascular expansion occurring during gestation.

The chick chorioallantoic membrane (CAM) is a common model for studying biological processes, but descriptions of the CAM circulatory system and especially experimental preparations of the CAM in shell-less eggs are both scant and controversial. We studied the CAM structure and the three-dimensional spatial configuration of the CAM vessels using five methods: in vivo stereoscopic observations, whole-mount preparations, histological sections, corrosion cast microinjection techniques, and the reconstruction of a three-dimensional wax model. Our findings show that the CAM consists of a superficial two-dimensional layer composed of a network of a very dense capillary mesh, floating over and enclosing a deeper three-dimensional space in which medium and larger free-floating vessels are seen to supply and drain the superficial layer. It is interesting to note that no tips or sprouts of blood vessels were observed during the development of the CAM vessels. In a shell-less egg preparation, the capillaries were found in the mesoderm layer of the CAM and not in or superficial to the ectoderm as is the case in the CAM of the intact egg which adheres to the shell membrane.

Ovarian hyperstimulation syndrome was induced in rabbit by administration of human menopausal gonadotropin (HMG) and human chorionic gonadotropin (HCG). In an attempt to establish whether serotonin plays a part in the induction of this syndrome, the hyperstimulated rabbits were divided into two groups and were administered known anti-serotonin drugs, cyproheptadine and methysergide, respectively. The group treated with cyproheptadine, a non-specific serotonin antagonist, exhibited significant acceleration in the regression of the syndrome. Methysergide, a specific serotonin antagonist, administered to the second group, neither prevented the occurrence of the syndrome nor accelerated its regression. The results of this work indicate that serotonin does not seem to be directly involved in the production of the ovarian hyperstimulation syndrome in rabbits.

The need for an adequate blood supply is of prime importance in successful skin grafting and in the take of keratinocyte cultures. Thus, the human uterine angiogenic factor (HUAF) extract, which induces neovascularization of the chorioallantoic membrane (CAM), was employed. The bioassay of HUAF was performed on an in vivo model of subdermally implanted collagen sponges and on sponges implanted into full skin thickness burn wounds in guinea-pigs. The HUAF extract was injected into the sponges every other day for 10 days. Each injection contained 10 micrograms decidual proteins with a total of 50 micrograms/sponge. The animals were sacrificed and the sponge together with the surrounding structures were extirpated, examined macro- and microscopically and by histological techniques. HUAF induced growth of blood vessels from the surrounding vascular bed into the implanted sponges. The angiogenesis was characterized by dense tortuous vessels with centripetal orientation. The control sponges exhibited only sporadic growth of blood vessels. This phenomenon repeated itself in the animals which were inflicted with burn wounds. The present study demonstrates that HUAF extract is also active on the in vivo model of experimental burns and wounds.

The effects of prolactin (PRL), alone and together with human chorionic gonadotropin (hCG), on steroidogenesis and cAMP accumulation in the preovulatory ovary were studied. Cultured granulosa cells obtained from large preovulatory follicles of pregnant mare serum gonadotropin (PMSG)-treated immature rats were used. The results indicated that PRL inhibited, in a dose-dependent manner, hCG-induced cAMP accumulation and 17 beta-estradiol (E2) secretion. When added to 0.4 IU/ml hCG (designated as 100% activity), 1, 10 and 100 ng/ml PRL decreased cAMP accumulation to 86, 64 and 59%, respectively, following 1 h incubation and to 87, 81 and 66% E2 secretion, respectively, following 48 h incubation. PRL alone failed to cause any significant change in cAMP or E2 concentrations. The inhibition of PRL was apparently not at the hCG receptor level, since a similar inhibitory effect was observed in prostaglandin E1 (PGE1)-induced cAMP accumulation. Nor was the inhibitory pathway of adenylate cyclase involved, since pertussis toxin--an inactivator of the Gi regulatory protein--failed to abolish the suppressive effect of PRL on hCG-induced cAMP accumulation. The phosphodiesterase (PDE) inhibitor, 3-isobutyl-1-methyl-xanthine, abolished the inhibitory effect of PRL on hCG- and PGE1-induced cAMP accumulation and on hCG-induced E2 secretion, indicating that PRL might be inhibiting cAMP accumulation and steroidogenesis in preovulatory granulosa cells by enhancement of PDE activity.

A plastic-cast technique was used to study the arterial vascular tree of the human uterus during pregnancy. The 3-dimensional models thus obtained revealed the organization, distribution, number, and diameter of the vessels. A vast ipsilateral and contralateral anastomotic network was evident throughout the uterus. The vascular structure reveals an increase in the size and degree of arborization of the arcuate arteries in and around the implantation site, which developed as a result of the increasing demand emanating from it.

Induction of a mild hyperstimulation syndrome in the female rat, and the possible effect of the induction on prolactin, were studied using light and electron microscopy as well as radioimmunoassay. Five days of human menopausal gonadotropin (HMG) treatment were followed by human chorionic gonadotropin (HCG) administration. Ovarian hyperstimulation was observed in all treated rats, as attested to by the weight increase and by histological studies. Ovaries were enlarged 3-4 times as compared with controls, and consisted mostly of corpora lutea follicular cysts and hemorrhagic cysts. Other visceral organs remained unaffected, except for slight edematous changes observed in the lungs of the treated rats. Electron microscopic examination of the hypophysis of the treated group tends to indicate increased synthesis and secretory activity of the prolactin cells. However, serum prolactin levels of treated and non-treated groups were not different. Augmented doses of the gonadotropins did not change the findings, and only mild grade 2 ovarian hyperstimulation, according to Schenker and Weinstein (1978), could be produced in the mature rat. Thus, the rat experimental model does not seem to be comparable with the situation in the human, and cannot be used to study the pathogenesis of the disorder in the human.

Microbiopsies of human Fallopian tube fimbrial epithelium were obtained during routine gynecological operations, and during pregnancy while undergoing hysterectomy in the first trimester or cesarean section in the third trimester. Specimens were studied under light and electron microscopes utilizing histo- and cytochemical techniques. Large juxtanuclear globules were demonstrated in the ciliary cell at all stages of the cycle. During the proliferative phase, these globules contained large lipid droplets and glycogen rosettes. The content of the globules in the early luteal phase was mostly amorphous electron-lucid material which failed to stain with PAS, alcian blue and the Millon reaction. However, reaction products of acid phosphatase as well as calcium precipitate were scattered in them as discrete granules. The same type of globule was found in early pregnancy. During late luteal phase and late pregnancy, the content of the globule was once again composed of lipids and glycogen. It seems that lipid and glycogen accumulate during the preovulatory phase, while depletion and dispersion of these substances occur in the periovulatory phase, probably due to increased energy utilization. Late luteal phase revealed subsequent replenishment of these reserves, as was also revealed in late pregnancy.

A method for inhibiting the adhesion of stone fragments in comminution is presented. Surfaces are coated with a molecular layer of so-called "excluding" polymer. In experimental tests, rat bladders were injected with comminuted urinary stone (phosphate or oxalate) in saline which contained small amounts of excluding polymer (0.1% hyaluronic acid or polyvinyl pyrrholidone). The urinary tract was then irrigated with fresh polymer solution and examined for residual stone. A ten-fold reduction in particle count was obtained, compared with irrigation with normal saline. The possible applications in percutaneous and extracorporeal lithotripsy are discussed. It seems that the "coagulum" and "exclusion" types of polymer irrigant might find complementary uses: the former for manual extraction of coarse particles and the latter for flushing out after fine comminution.

The human Fallopian tube fimbrial epithelium was histo- and cytochemically studied in the first and third trimesters of pregnancy. In the first trimester morphometric measurements confirmed a decrease in cell size and a reduction in ciliary cell count. The ciliary cell population was surprisingly glycogen-poor, and some of the cell contained, among normal organelles, lipid accumulation, lamellated bodies and nuclei with slightly condensed chromatin. Some had defective cilia and basal bodies. Large globules, containing few lipid droplets and glycogen rosettes were demonstrated. The bulk of the globules, however, contained an unidentified electron-lucid substance, negative to PAS, Alcian blue, OsO4 and the Millon reactions, but positive to acid phosphatase reaction, the products of which appeared dispersed. The nonciliary cell population was heterogeneous; some appeared almost devoid of microvilli and had dilated RER and SER cisternae, lamellated bodies and condensed nuclear chromatin. In some cells a single centrally located cilium was noticed. Other nonciliary cells had extensive microvilli and appeared less atrophied. In the third trimester morphometric, as well as cytochemical methods indicated that the regressive process was beginning to reverse itself. Alkaline-phosphatase reaction products appeared along the apical and lateral membranes of the nonciliary cell. Acid-phosphatase reaction products depicted the lysosomes, present in both types of cells.

Ultrastructural and cytochemical methods were utilized to study the human Fallopian tube fimbrial epithelium during the different stages of the menstrual cycle. Alkaline phosphatase reaction product was located along the apical and lateral plasma membranes of the secretory cells only, regardless of the stage of the cycle. The ciliated cells were almost devoid of any reaction product at all stages of the cycle. Acid phosphatase reaction product depicted the lysosomes. These appeared as electron-dense bodies, of almost equal numbers in the ciliated and the secretory cells at all stages of the cycle. Thus the number of lysosomes did not vary appreciable during the different stages of the menstrual cycle. Many lipid droplets were found in both cells; these were rimmed by acid phosphatase reaction product, and some were partially enveloped by electron-dense bodies containing acid phosphatase deposits. Acid phosphatase deposits were also found on the inner face of Golgi vesicles.

The effects of prolactin (PRL), alone and as a modulator of human chorionic gonadotropin (hCG) action, on steroidogenesis and cAMP accumulation in rat luteal cell cultures were examined. Cultured rat luteal cells were prepared from immature rats primed with pregnant mare serum gonadotropin and hCG. In vitro treatment was performed with 0.1 and 0.2 IU/ml hCG and 1-100 ng/ml PRL. Cultures were incubated for 48 h for evaluation of progesterone (P4) secretion and for 1 h for measurement of cAMP accumulation. The same doses of hormones and incubation periods were also used in preovulatory rat granulosa cell cultures and found to cause a significant, dose-dependent inhibition in estradiol, P4 and cAMP accumulation. In luteal cell cultures, on the other hand, P4 secretion was significantly elevated, in a dose-dependent manner, by PRL. Moreover, identical doses of PRL caused a significant, dose-dependent stimulation of cAMP accumulation. Basal levels of P4 were also significantly elevated by PRL alone, but no such stimulation by PRL was detected in basal levels of cAMP. Addition of the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, increased the stimulation of P4 and cAMP by hCG + PRL in a manner dependent on PRL concentrations. The overall data therefore demonstrate divergent effects of PRL on cAMP accumulation and steroidogenesis in the ovary: inhibitory in the preovulatory and stimulatory in the postovulatory state. Moreover, these findings suggest a possible common mechanism linking the effects of PRL before and after ovulation: inhibition of cAMP accumulation via enhanced breakdown of the nucleotide.

Mitogenic preparations of nonviable lysed cells and purified membranes of Mycoplasma pulmonis induced interstitial pneumonia and tracheitis after intranasal administration to pathogen-free rats. The pneumonia, characterized by peribronchial, perivascular, and alveolar wall infiltration by lymphocytes, was indistinguishable from that produced by viable M. pulmonis. Both pathologic and mitogenic effects were significanlty reduced by prior treatment of membranes with heat or proteolytic enzyme. Intranasal administration of the thymus-derived-cell mitogen, concanavalin A, produced interstitial pneumonia but not tracheitis. These results indicate a correlation of mitogenicity and pathogenicity and suggest that activation of thymus-derived lymphocytes is the major cause of the pneumonia resulting from infections with M. pulmonis.

Most of the morphological changes which occur during the different phases of the reproductive cycle are hormone-dependent. Thus, the aim of this work was to demonstrate the fluctuations in the availability of steroid binding sites in the ampullary segment of the tube during the menstrual cycle. Specimens were obtained during routine gynecological operations and were prepared for histological examination. In addition, determinations of specific binding of estradiol and progesterone (specific activity was about 100 Ci/mmol) were made in soluene-dissolved tissue of ampullary mucosa. Autoradiographic grain counts of tissues which were incubated with [3H]estradiol and [3H]progesterone were performed on histological sections. The epithelial cell binding capacity was higher than that of the cells in the underlying lamina propria and this was found throughout the cycle. Estrogen and progesterone binding capacity was higher during the proliferative phase and decreased markedly during the secretory phase. Tissue binding of steroids appears to be inversely related to serum levels, which are lower in the proliferative phase and rise during the secretory phase.

Ectopic pregnancy specimens of 6 human Fallopian tube fimbrial epithelia were studied utilizing ultrastructural, cytochemical and morphometric methods. The observations were compared with those made on 12 intrauterine pregnancy specimens. The morphometric measurements indicate earlier onset of atrophic changes in ectopic pregnancy. Deciliation in Ectopic pregnancy preceded that observed in intrauterine pregnancy. Cell height also decreased earlier in ectopic pregnancy than in intrauterine pregnancy. In addition, during the 1st trimester, ultrastructural localization of calcium revealed that, in ectopic pregnancy, the mitochondrial calcium, which was observed in intrauterine specimens, was shifted into the cytoplasmic compartment of the ciliary cell. Consequently, cytoplasmic calcium was found in the ectopic specimens and was less evident in the intrauterine specimens. Thus, a local atrophic effect is evident in ectopic pregnancy, which appears earlier than the atrophic process found in intrauterine pregnancy.

Microbiopsies of 38 specimens of human Fallopian tube fimbrial epithelium obtained during hysterosalpingectomy operations were processed for electron microscopic observation. Calcium localization was demonstrated utilizing the pyroantimonate-osminium-tetroxide technique. Calcium deposits were consistently observed as scattered granules in the nuclei, nucleoli, and cell membranes of secretory and ciliary cells regardless of the stage of the menstrual cycle. On the other hand, mitochondrial-bound calcium was observed in the ciliary cells during the preovulatory phase, coinciding with relatively high estrogen levels, and was not found during the peri- and postovulatory phases of the menstrual cycle. It appears that there is cellular calcium which is relatively stable and is probably part of the structural components of the cells. The ciliary cell mitochondrial calcium appears to be more labile and is therefore available for mobilization and utilization in the metabolic processes associated with the periovulatory phase.

Morphological changes were correlated with biochemical data induced by prolactin (PRL) in cultured rat granulosa cells from large preovulatory follicles. Biochemical results indicated that PRL exerted a significant dose-dependent inhibition in gonadotrophin-induced secretion of progesterone and 17 beta-oestradiol. PRL alone failed to affect basal steroidogenic secretion. In parallel morphological experiments, using phase-contrast microscopy, untreated and 100 ng/ml PRL-treated cells appeared as a monolayer of flattened, fibroblast-like cells. Upon exposure to 0.4 IU/ml human chorionic gonadotrophin (hCG), aggregates of rounded, epithelioid-shaped cells were formed. The addition of PRL to hCG in the same doses minimized the changes induced by hCG. Similarly, electron microscopy of untreated and PRL-treated cultures revealed flat cells devoid of microvilli, with evenly dispersed microfilaments. The addition of hCG caused rounding of the cells and was accompanied by the appearance of microvilli and by pronounced steroid-producing organelles. Bundles of microfilaments were noted at the cell periphery. PRL added to hCG caused a reduction of the hCG effects, and the cell morphology was intermediate to that seen in untreated and hCG-treated cultures. The finding that PRL can prevent or minimize morphological changes caused by hCG in rat cultured granulosa cells correlates with the biochemical changes induced by PRL, and supports the concept that PRL is a modulator of gonadotrophic action in the ovary.

Ultrastructural and cytochemical features of 100 specimens of human Fallopian tube fimbrial epithelium were studied to determine whether they were dependent on or independent of the ovarian hormones. Characteristics which were independent of hormonal control included the presence of alkaline phosphatase reaction product along the apical cell membrane of the secretory cell and its absence in the ciliary cell. Acid phosphatase reaction product was found in ciliary and secretory cell lysosomes in similar numbers and calcium deposits were observed in ciliary and secretory cell nuclei, nucleoli and cell membranes. On the other hand, characteristics dependent on stages of the cycle included the presence of mitochondrial-bound calcium during the preovulatory phase and its depletion during the periovulatory phase. The same sequence was noticed in the content of the large ciliary cell perinuclear globules, which displayed accumulations of glycogen and lipid in preovulatory phase and their depletion during the periovulatory phase. It seems that during the periovulatory phase the increased metabolic activity of the cells is accompanied by utilization of some of the material stored within the cells.

The mitogenicity and pathogenicity of Mycoplasma pulmonis were compared in two rat strains. Both the mitogenic and the pathologic effects induced by M. pulmonis membranes were more severe in Lewis rats than in Hooded rats, and were dependent on the mitogen doses used. It was concluded that the severity of lung lesions induced by M. pulmonis membranes correlated with the degree of mitogenic responses of the different rat strains to this organism. The roles of T- and B-cell mitogens in induction of pneumonia were studied in Hooded rats treated intranasally with either the T-cell mitogen concanavalin A or with M. neurolyticum membranes which stimulate the B-cell populations, or with both concanavalin A and M. neurolyticum. Results clearly showed that the individual B- and T-cell mitogens affected the lungs of treated animals. Nevertheless, the mitogenic co-stimulation of both B and T lymphocytes in rat lungs was necessary to obtain maximal development of interstitial lymphocytic pneumonia.

Meshed human dermis allografts containing human uterine angiogenic factor were implanted over full thickness surgical skin wounds in rats. Enhanced angiogenesis of the wound bed, three times more than that observed in control grafts, was found in experimental grafts at 3 days postimplantation. This was accompanied by a greater amount of granulation tissue formation, enhanced granulation tissue penetration into the dermal grafts and accelerated incorporation of the grafts. Angiogenesis of the wound bed regressed to control levels at 7 days postimplantation. The growth and penetration of the granulation tissue and the accelerated incorporation of the dermis grafts, however, continued ahead in the experimental grafts while necrosis appeared in the control grafts. The relevance of these results for acceleration of wound healing and improvement of the wound bed for subsequent application of cultured epidermal grafts is discussed. This method may be extended to the treatment of ulcers and burns.