The prevention of osteoporotic progression by means of steroid loaded TCPL drug delivery systems.

Numerous studies have suggested that there is a link between the age-related decreases in Estradiol and adrenal androgens, and the subsequent development of senile osteoporosis. The specific objectives of this investigation were: 1) to deliver Dehydroepiandrosterone (DHEA), Diosgenin (DG), and Estradiol (E) at sustained levels by Tri-Calcium Phosphate Lysine drug delivery systems (TCPL), and 2) to study the effects of the sustained delivery of DHEA, DG, and E on the bone turnover of adult female rats following withdrawal of the endogenous hormonal milieu by means of ovariectomization (OVX). In this study, 20 female Sprague-Dawley rats were randomly divided into five groups containing four rats per group. The rats in Group 1 served as intact controls. Animals in groups 2-5 were ovariectomized, and groups 3-5 were implanted immediately with TCPL drug delivery capsules containing DHEA, DG, and E, respectively. Group 2 served as the SHAM (OVX only) group. At the end of 33 days post implantation, the vital organs, reproductive organs, and femurs were collected and evaluated. Bone histomorphometric analyses as well as mechanical strength testing was performed. Data obtained from this study demonstrated that body weights were increased in all OVX animals, and that E replacement resulted in body weights that were not significantly different from intact controls. No differences were seen in the wet weights of any vital organs. However, a decrease in the weights of the cervix and oviducts were evident in all ovariectomized groups, with the exception of the E group. Thirty-three days following OVX, the OVX-only group exhibited an increased inner medullary area, decreased thickness of the cortical layer of bone, and decreased mechanical strength. The group treated with DHEA and the group treated with E were shown to maintain both the medullary area and the cortical thickness (as compared to the intact control group). The three point bending test of the femora showed that OVX-only induced a slight decline in mechanical strength, and that DHEA and DG, but not E, showed increases in mechanical strength. Results of this experiment suggest that DHEA and E may reduce bone remodeling as evidenced by the reduction in the medullary area, and that DHEA may possibly be used in postmenopausal patients to reduce osteoporotic progression.

The effects of thyroid and reproductive hormones on the viability of human buccal epithelium.

Recent findings have shown that estrogen receptors were localized in stromal cells in the oral cavity. The role of physiological doses of growth promoting hormones such as thyroid stimulating hormone (TSH) and estrogen have not exclusively studied. This investigation identifies the cytology and biochemical response of buccal epithelial in the presence of estrogen and TSH at 24, 48, and 72 hr of incubation. The cells were obtained from pre, postmenopausal women and men. The cells were seeded at a density of 10,000 cells per well and aseptic techniques, morphological evaluation, biochemical analysis (MDA), and data analysis were performed following standard lab protocols. Results of this investigation revealed that: (i) there was an initial increase in total protein observed in cells treated with TSH in comparison to the control and estrogen treated cells. This trend continued for 48 hours, and no statistical differences were observed at the 72 hour phase, (ii) there was an initial decrease in MDA levels in cells treated with TSH, however, cellular damage was evident at 48 and 72 hours phases in comparison to estrogen and control groups, (iii) morphological evaluation demonstrated that there were structural changes associated with TSH treatment. These include aggregates, nuclear shriveling and lack of cellular boundaries. Estrogen treatment showed no structural alteration throughout the experiment.

The effect of thyroid stimulating hormone on the proliferation and viability of HEP-2 laryngeal cells in culture.

The objective of this experiment was to study the effect of TSH on Hep-2 cells. A total of sixteen tubes (5 x 10(4) cells per tube) were divided into four equal groups (media alone (control), serum containing 0 TSH, 10.3 microliters/ml TSH, and 49 microliters/ml TSH). The supernatants and cells were collected at 24, 48, and 72 hours after incubation. The result show that TSH caused an increase in cell number after 24 hours in comparison to control media alone. Analysis of supernatants for cellular damage showed an increased pattern in MDA levels in serum exposed cells at 24, 48, and 72 hours. In contrast, MDA levels in TSH treated cells were similar at 24, 48, and 72 hours. The levels obtained at 48 and 72 hours were statistically (P < 0.05) lower than those obtained for control and serum treated or 0 TSH group. This observation suggests that TSH could provide a protective measure against membrane lipid peroxidation. Morphological evaluation of the cells at 24, 48, and 72 hours, suggests that TSH exposure did induce noticeable cellular injury and most adaptive responses observed were shape changes (round up), cellular detachment, and hyperchromatic nuclei (decrease in cell number).

Morphometric analysis of the adrenal compartments exposed to sustained delivery of androgens.

The role of reproductive and adrenal steroid hormones on the structural and functional capacity of adrenal tissue has not been well investigated. The objective of this investigation was to morphometrically evaluate the effect of testosterone (T), dihydrotestosterone (DHT), and androstenedione (AED) given in a sustained manner, by tricalcium phosphate lysine (TCPL) ceramic capsules, on the adrenals of adult male rats. Sixteen adult male rats were randomly divided into four equal groups. Groups II, III, and I were implanted with TCPL capsules loaded with AED, T, and DHT, respectively. Group IV animals were not implanted, and thus served as the control group. At the end of ninety days post-implantation, the animals were euthanized using standard aseptic surgical techniques. The adrenal glands were harvested and stored in 10% formalin. The tissues were processed, embedded, sectioned and stained with H & E using standard laboratory procedure. Random sections of control and experimental tissues were utilized for morphometric analysis by using Image Pro digital analysis techniques. Data collected were analyzed by means of ANOVA (p < 0.05). Results of this study revealed. (1) There was an increase in the total areas of T treated animals in comparison to the control and other experimental groups, (2) the total lengths of each hormonally treated tissue showed an increase in size of DHT treated tissue verses control, but differences of T and AED compared with control remained insignificant, (3) upon analysis of the zona glomerulosa (Z1) and zona fasciculata (Z2) the data demonstrated a significant increase in animals treated with DHT and AED in comparison to control and T treated animals, (4) finally, statistical analysis performed on measurements of the zona reticularies (Z3) indicated notable increases only in the AED exposed animals. The changes in size of the various tissues may be warranted due to reactions of the steroid hormones with different surface receptors in different layers. However, further investigations are needed, especially longer duration of treatment, to fully hypothesize the exact mechanisms of these hormones on the adrenal glands.

Development of a model for bioresorbability of bioceramic microparticles.

Over the past decade, numerous studies have implicated wear particle debris, generated by everyday wear or ceramic implants, as the essential contributors to implant failure [3,4,5,6,7]. We investigated the interactions of ceramic particles (TCP and HA) on cell viability, cell damage, and respiratory burst in a macrophage cell line (RAW 264.7) for a duration of 24, 48, and 72 hours. The results show that cellular protein levels ranged between 0.20-0.3 mg/ml for the duration of the experiment, and there was no difference detected between or among the groups (P < 0.05). Differences in MDA levels (the amount of lipid peroxides formed at the cellular membrane) were not detected at the early time points (24-48 hours). However, with time in culture there is an increase in MDA levels in all groups. There were no differences detected in nitric oxide production between TCP and the control at all phases. The amount of nitric oxide produced by cells treated with Ha decreased slightly at 24 hours. In contrast, at 48 and 72 hours there was a significant increase when compared to the control. Morphological evaluation revealed that cells treated with TCP contained irregular membrane boundaries, and appeared to have cytoplasmic extensions. Many of the macrophages consisted of large vacuoles containing TCP particles. Cells treated with HA also contained irregular cytoplasmic borders with an increase in vacuolization. At 48 hours, the morphology of all groups remained the same, and at 72 hours, many of the cells treated with TCP and HA began to lyse. Overall, RAW 264.7 macrophage cells are capable of ingesting particles of TCP and HA, and after three days in culture the cells begin to show the effects of increased retention of ceramic particles. The results of this investigation can be used as a model for the assessment of the resorbability rate of implantable devices.

An evaluation of infectious diseases in cervicovaginal smears from patients with atypical cells of undetermined significance.

Despite all studies of infectious disease of the female genital tract, there have been a few studies of the many different types of infectious organisms on Pap smears that contain abnormal cells. The purpose of this study is to evaluate the significance of infectious organisms in women with a cytologic diagnosis of atypical squamous cells of undetermined significance (ASCUS). This study utilizes 398 cervicovaginal smears and concurrent cervical biopsies (test and control cases) obtained from the Department of Health cases from January to December 1993 and 1995. The cases were randomized, assigned a study number and reviewed for the presence of infectious organisms. The age range for the patients consisted of 15 to 62 for the ASCUS test group (mean 24) and 13 to 64 for the matched control group (mean 24). The results of the microscopic review were entered into SPSS for evaluation. Infectious organisms were identified in 101 (25.4%) of the test and control cases. The type of organisms identified in the ASCUS and matched control cases were normal vaginal flora 42 (21.2) and 48 (24.0%), Actinomyces one (.3%) case, Candida albicans in 2 (1%) and 4 (2%), Trichomonas vaginalis in 10 (5.1%) and 9 (4.5%). Herpes virus effect was not identified in the test cases but was identified in 2 of the control cases. The cytopathic effect of human papilloma virus was identified in 68 (34.3) and 3 (1.5%) of the test cases. Data collected in this study indicate that infectious organisms, other than the effect of the human papilloma virus, are not as common in the ASCUS cases as the matched control cases. Human papilloma virus effect was more commonly associated with the squamous intraepithelial lesions.

Cytohistological correlation of abnormal cervical lesions in Native American females in Mississippi.

Earlier studies have reported that cervical cancer mortality and incidence rates in Native American women are one of the highest in the United States. In recent years, the rate of abnormal cervical lesions in this group has been a public health concern. The objective of this study was to evaluate a cytohistologic correlation of abnormal cervical lesions in Native American women in Mississippi. The cytohistologic results from 77 Native American women with an abnormal diagnosis were reported to the University of Mississippi Medical Center during 1993-1996. The cases were divided into four groups; which included 7(9%) atypical squamous cells of undetermined significance (ASCUS), 51(66%) low-grade squamous intraepithelial lesions (LGSIL), 19(25%) high-grade squamous intraepithelial lesions (HGSIL), and no invasive carcinomas. Out of the study cases, 57 women presented with an abnormal gynecological history. The mean age was 29.97 years with a range of 14 to 87. The results were tallied and evaluated using SPSS. Only, 57 cases had a histologic evaluation, with an overall correlation rate of 84% (48/57). The histopathologic diagnosis confirmed 12 negative, 26 CIN I, 17 CIN II/CIN III, and 2 invasive carcinomas. The results of this study conclude that cervical intraepithelial lesions are significant among Native American women in Mississippi. Fifty-six (98%) of the patients with histologic evaluations presented with an abnormal gynecologic history; therefore, education and routine cytologic screening is imperative for the detection of cervical cancer in high-risk groups.

The use of estrogen, DHEA, and diosgenin in a sustained delivery setting as a novel treatment approach for osteoporosis in the ovariectomized adult rat model.

It is well established that the pattern of bone loss from the cortex in osteoporotic bone begins from the endosteal surface of the cortex, where there is enlargement of the medullary canal at the expense of the inner cortex. Bone loss does not occur at the periosteal surface. The objective of the following study was to induce osteoporosis in female rats by ovariectomy, followed by treatment with sustained delivery of Diosgenin (DG), dehydroepiandrosterone (DHEA), or estrogen (E) after clinical signs of osteoporosis. Female Sprague Dawley rats were divided randomly into five groups containing four rats/group. Rats comprising group 1 were left intact and served as a control group. Animals in groups 2-5 were ovariectomized (OVX) and, after a 14 day delay to allow for induction of osteoporosis, were implanted with TCPL capsules containing DG, DHEA, and E, respectively. The experiment was ceased after 33 days of treatment, at which time the vital and reproductive organs for each group were collected, weighed, and analyzed histomorphometrically for differences. Further analysis of the progression of osteoporosis in the experimental animals was obtained by performing x-ray analysis of each group on a semi-weekly basis. By collecting and analyzing the femurs from each animal, we were also able to obtain important information about the histologic changes associated with osteoporosis (left femur), as well as data regarding the effects of osteoporosis on the mechanical strength of bone via three point bending analysis (right femur). The data generated by this study revealed important information as to the efficacy and safety of the alternative treatments DHEA, E, and DG for osteoporosis. First, histomorphometric analysis revealed that treatment with DHEA, E, and DG reduced the endosteal perimeter and cortical area to values very similar to controls (intact). Second, results of the bending stress and modulus in OVX and treated animals were not statistically different from the intact control animals, which suggests that the material properties of the bone were unaltered. Third, there is an increase in total body weight associated with OVX that is reduced to control levels after replacement therapy. Finally, OVX also resulted in reproductive tissue atrophy, which was reversed by all three of the treatment regimens in this study. These data suggest that bone loss after OVX can be significantly reduced by supplementation with sustained levels of DHEA, E, and DG without jeopardizing other body organs.

The physiological response associated with large particles of TCP, HA, and ALCAP implants using raw 264.7 cells.

Ceramic materials were introduced as implantable materials in the early 1960s. Currently, there are numerous types of ceramic materials under evaluation as ideal implant devices. The inflammatory responses generated by various calcium phosphate-based ceramics implants have not been clearly evaluated. The current investigation evaluates the response of RAW 264.7 macrophage cells to large particle size (> 38 microns) of hydroxyapatite (HA), tricalcium phosphate (TCP), and aluminum calcium phosphate (ALCAP). RAW 264.7 cells were cultured in a 24 well plate at a density of 1.5 x 10 cells per well. The plates were divided into seven equal groups (n = 6) (control, HA, TCP, ALCAP, ops HA, ops TCP, ops ALCAP). The total protein, maliondialdehyde (MDA), nitric oxide, and cell counts were measured using established lab protocols at 24, 48 and 72 hours of incubation. Cells grown on coverslips were used to evaluate morphological features. The data obtained from this investigation suggested that the nonopsonized particles of ALCAP and TCP materials exhibited an increase in cell number at both the 48 and 72 hour phases. However, a decrease was seen in cell number in all three opsonized groups at 48 and 72 hour phases when compared to the control. As for MDA levels in the opsonized group, all treatments showed an initial increase. Nitric oxide production increased in TCP and ALCAP at the 48-hour phase. Morphological evaluation revealed that upon the exposure of the three different type of ceramic, the cells appeared to be biocompatible.

Morphological and biochemical evaluation of RAW 264.7 cells exposed to polychlorinated biphenyl.

The effects of PCBs exposure, at low doses ranging between 10-100 mg PCB (Arclor-1254), on the viability of RAW 264.7 macrophage cells after 24, 48, and 72 hours of incubation was investigated. Furthermore, this study was designed to determine the interrelationship between PCBs and estrogen (E, 10 mg) and its role in the viability of RAW cells. Macrophages were cultured and plated in 24 well-plates according to standard protocols. The wells were divided into eight groups (n = 4 wells/group, 1 x 10(4) cells/well). The cells in groups 1-3 were treated with vehicle (serum-control for estrogen, DMSO-control for PCB, and media alone, respectively). Cells in groups 4 and 5 were treated with low (10 mg) and high (100 mg) doses of PCB. Cells in group 6 were treated with E, and cells in groups 7 and 8 were treated with low dose PCB + E and high dose PCB + E, respectively. Cell viability and damage (Malianodialdehyde, MDA level) were determined after 24, 48, and 72 hours, as well as, cell morphology. The results of this study showed that low and high doses of PCB depressed cell number by 52%. Estrogen treatment caused no effect on cell number in comparison to cells treated with serum alone. Cell number in response to E and low and high doses of PCB decreased cell number by 50%. Similar results were also observed at 48 hour time phase. In contrast, at the 72 hour phase, no significant changes were observed for cell count. Morphological evaluation of the cells revealed healthy spindle shaped multinucleated cells in the control group but, groups exposed to PCB induced morphological changes that included: cells became small, round, and increased evidence of cellular injury and debris. Estrogen treatment did not show changes from the control group. However E and PCB treatments caused the cells to become round, tightly compact nuclei with evidence of cell fragmentation. The results of this investigation showed that exposure to either 10 or 100 mg of PCB had detrimental effects on the RAW 264.7 macrophage cells as early as 24 hours. Combination treatment with E didn't provide any protective measures to the viability of RAW cells exposed to PCBs.

The effects of inositol hexaphosphate on the inflammatory response in transformed RAW 264.7 macrophages.

Inositol hexaphosphate (IP6) has received much attention for its role in interfering with tumor progression and slowing the metastasis of neoplastic cells. However, there is little information regarding the antioxidant properties of IP6 or its ability to enhance the natural disease resistance of the body. The specific objectives of this experiment were to investigate the effects that IP6 might have on the proliferation and viability of RAW 264.7 transformed macrophages and to morphologically and biochemically investigate the role of IP6 as a free radical scavenger. Transformed RAW macrophages were obtained from the American Type Culture Collection (Rockville, MD) and maintained in sterile media (RPMI) supplemented with 10% fetal bovine serum and 1% antibiotics and antimycotics. The cells were plated on to 24 well plates at a density of 1 x 10(5) cells/well. The cells were divided into five groups of four wells per group per phase (24, 48, and 72 hours). Cells in Group I were treated with media alone and served as controls. Cells in Group II were treated with lipopolysaccharide (LPS) only. Cells in groups III, IV, and V were treated with 1000 microliters of IP6 + LPS, 500 microliters of IP6 + LPS, and 100 microliters of IP6 + LPS, respectively. Cell numbers, as well as, morphology, MDA, and protein were determined at the end of 24, 48, and 72 hours. Data obtained from this investigation revealed that the rate of cell proliferation was totally dependent on the dose of IP6. At 24 and 48 hours and upon the exposure of high dose of IP6 the mitotic ability of the cells was higher (p < 0.05) than the rate at the 72 hour phase. Morphological evaluation of cells at all three phases revealed that there were significant changes in the architecture of cells upon the exposure of IP6 compared to the control group. The results of this study suggest that IP6 may have had an excitatory effect on the inflammatory cell secretions and this phenomenon was found to be dose dependent.

The role of the route of administration of poly-chlorinated biphenyls (PCBs) on the reproductive and vital organs of adult female rats.

Studies have shown that high doses of Polychlorinated biphenyls (PCBs) given by conventional methods (orally or injections) have adverse effects on the reproductive and vital organs of adult female rats. However, there has not been documentation as to the effects of PCBs on adult female rats by means of a sustained delivery system. The specific objectives of this study were: (1) to investigate the effects of sustained delivery (TCPL ceramics) of PCB versus conventional mode of administration (injection) on the reproductive and vital organs of the adult female rat, (2) to evaluate the role that PCB that might have on the estrus events of adult female rats upon the exposure by sustained delivery (TCPL ceramics) and conventional mode (injection), and (3) to histopathologically evaluate the effect that PCB might have on the ovarian and accessory organs upon the sustained delivery for 21 days. A total of 10 adult female rats (BW 270-300 gm) were randomly divided into three groups. Group 1 (n = 3) served as the control, group 2 (n = 4) was injected intramuscularly every other day with Aroclor 1254 (0.1 cc), and each rat in-group 3 (n = 3) was implanted with TCPL capsules (5 mg of 2,3,3',4,5-Pentachlorobiphenyl). Aseptic surgical techniques were performed throughout the experiment. Blood (1 cc) was collected biweekly for biochemical analysis, and body weights were recorded as well. Pap smears were taken daily at approximately the same time for 25 days, and two slides were made for each pap taken (1 pap stain, 1 Diff Quik). At the end of 21 days post-implantation, all control and experimental animals were sacrificed following standard lab procedures (overdose of Halothane). The reproductive and vital organs were collected, weighed, fixed, embedded, sectioned, and stained (H&E) for histological evaluations. Data obtained from this investigation suggest the following: (1) TCPL devices were able to deliver PCB at sustained levels for 21 days, (2) regardless of the route of PCB administration no significant change was observed in total body weight compared to the controls, (3) conventional administration of PCB resulted in a remarkable changes in the fallopian tubes compared to control and sustained delivery implanted animals, (4) there were no obvious change was observed in the phases of estrus cycles upon the exposure of PCB, and (4) histopathological evaluation of spleen, kidneys, heart, adrenals, ovaries, uterus, and cervix tissues exposed to PCB did not reveal any significant changes compared to the intact group.

TCPL drug delivery system: the effects of synthetic DHEA and Diosgenin using an ovariectomized rat model.

Dehydroepiandrosterone (DHEA) has been shown in numerous studies to exhibit a host of benefits at the vital and reproductive organ levels. However, the use of naturally occurring DHEA is hindered by its inability to survive the first-pass metabolic process of the liver. One possible alternative means that deserves consideration is the administration of DHEA's precursor, namely, Diosgenin (DG). The specific objectives of this investigation were: 1) to deliver DHEA and DG at sustained levels by Tri-Calcium Phosphate Lysine (TCPL) drug delivery systems using ovariectomized (OVX) adult rats as a model, and 2) to evaluate the biochemical and histopathological changes associated with the sustained delivery of DHEA and DG. A total of 30 adult female rats were used in this investigation. The animals were further divided into 8 groups. Groups 1 and 2 animals served as intact control groups while each rat in groups 3-8 was ovariectomized (sham (33), n = 3 [group 3], sham (47), n = 4 [group 4]). Groups 5 and 6 were implanted with DHEA (group 5) and DG (group 6) loaded TCPL capsules immediately following the OVX procedure. Groups 7 and 8 were implanted with DHEA (group 7) and DG (group 8) loaded capsules 14 days following OVX. Surgical aseptic technique was employed according to standard laboratory protocols. Maliondialdehyde (MDA) and hormonal levels were measured from serum, collected semi-weekly, during the entire investigation (for 47 days) and X-rays were performed weekly. Pap smears were collected daily for 47 days to assess endometrial changes associated with DHEA and DG treatment. Following sacrifice (at day 33 for groups 1, 3, 5, and 6 and at day 47 for groups 2, 4, 7, and 8), routine H and E staining was conducted for histopathological evaluation of the reproductive and vital organs. Results of this investigation demonstrated: 1) OVX resulted in an increase in total body weight, and the use of DHEA and DG returned the body weight to near normal levels as compared to the intact control groups, 2) TCPL capsules delivered DHEA and DG at a sustained level during the 47 day study, 3) serum levels of MDA are as follows: DG > DHEA = OVX > control for the 33 day phase and OVX > DG > DHEA > control for the 47 day phase, 4) no significant changes were observed in total wet weights, as well as the morphology of the spleen, kidney, adrenal, heart, liver, and lung tissues, 5) OVX resulted in an atrophy and non-keratinization trend in the reproductive tissues, and sustained delivery of DHEA and DG showed no remarkable change in these tissues, 6) the use of sustained delivery of DHEA and DG resulted in higher weights of uteri compared to the OVX group. In conclusion, this study provided more information regarding the interrelationship between DHEA and DG, and the physiological responses encountered when they are administered continuously using the adult OVX rat as a model.

Assessment of endometrial function during sustained delivery of estradiol and estradiol plus progesterone in ovariectomized rats.

Previous studies conducted in our laboratories have documented that tricalcium phosphate lysine (TCPL) delivery system can be utilized to deliver estrogen (E) and combination of estrogen with progesterone (E + P) at sustained levels for long duration in intact rats. The specific aim of this investigation was to attempt to reverse the endometrial changes resulted from post-ovariectomy by exogenously delivering sustained levels of E and E + P. A total of 27 adult female rats were used in this study. The animals were randomly divided into seven different groups: groups 1 animals served as intact control group, groups 2 and 6 animals were ovariectomized, 3 and 7 were ovariectomized (OVX) and implanted with TCPL loaded with E (20 mg loaded TCPL), and group 4 was OVX and implanted with E (TCPL, 20 mg) + P (TCPL, 40 mg.) treatment. Group 5 animals served, as intact control group. Blood samples were collected biweekly for 47 days. Vaginal smears were taken and screened daily during the entire investigation. Histopathological evaluations were conducted on reproductive as well as vital organs (H & E). Data obtained from this investigation suggest the following: (I) OVX resulted in an increase in total body weight, however E and E + P maintained the body weights at prior ovariectomy level, (II) sustained delivery of E resulted in maturation of vaginal epithelium and the smears exhibited the estrus at the end of 72 hours post implantation and continued this trend for the duration of the study, (III) E + P treatment induced no estrus and the epithelial changes resembled the OVX group, and (IV) E and E + P treatment resulted in a significant different (P < 0.05) in MDA levels compared to OVX and intact control groups. Results of this investigation conclude that sustained delivery of E and E + P by TCPL can be utilized to maintain the normal function of the reproductive organs and could serve as an efficient and safe therapy in estrogen deficient patients.

Cytomorphological assessment of benign and malignant dense hyperchromatic groups in cervicovaginal smears.

Dense hyperchromatic cell groups are considered common diagnostic problems in cytopathological evaluations. Clusters of cells with scant cytoplasm and dark nuclei represent the morphological features of dense hyperchromatic cells. Cytological evaluations of the dense hyperchromatic groups in cervicovaginal smear results in high rates of false-positive or false negative diagnosis. The key element is to differentiate among the dense hyperchromatic groups and to appropriately classify, based on strict morphologic criteria. The objective of this study was to evaluate the cytomorphology of benign and malignant dense hyperchromatic groups in cervicovaginal smears reported at the University of Mississippi Medical Center. Six distinct types of dense hyperchromatic groups were selected (forty-eight cervicovaginal smears) to represent all of the entities. The cases were divided into; group 1; atrophic pattern (n = 9), group 2; endocervical cells (n = 9); group 3, endometrial cells (n = 10), group 4; high-grade squamous lesions (HSIL)(n = 10), group 5; squamous cell carcinoma (n = 5), and group 6; endometrial adenocarcinoma (n = 5). Light microscope techniques were used to evaluate several parameters--such as, background, arrangement, and chromatin pattern. ImagePro digital analysis computer software (at x40 magnification) was used to quantify and evaluate the nuclear area and nuclear to cytoplasm ratio. Data obtained from this investigation suggest that there were significant differences observed in the total nuclear areas among all groups. In conclusion, cytomorphometric analysis can be utilized as an ideal diagnostic tool in differentiating between the ambiguous or suspicious groups of dense hyperchromatic cells. Ultimately, this diagnostic tool can minimize the rate of false-positive or false-negative diagnosis resulting in better cytologic evaluations and patient management.

Cytohistologic correlation in patients with clinical symptoms of postmenopausal bleeding.

Today, the life expectancy for women is longer; therefore, many will likely experience the postmenopausal period (termination of fertility and menstrual bleeding). Uterine bleeding after this period is a sign of pathologic condition. The specific objective of this project was to evaluate the cytohistologic findings in women with postmenopausal bleeding (PMB) and to determine the presence of any significant pathologic lesions. Cytohistologic correlations from 66 patients attained in 1993 from the University of Mississippi Medical Center were evaluated. The population evaluated were divided into three groups: (control group 1) dysfunctional uterine bleeding (DUB), (control group 2) postmenopausal (PMP), and (test group 3) the group of women with postmenopausal bleeding. The DUB and PMP age-matched controls (n = 12, mean age 51 +/- 5 and 57 +/- 5 years) were randomly selected, and correlated with the actual group being tested (54 PMB, mean age 57 years). The distribution among the 54 PMB women evaluated were 69% (37/54) black, and 31% (17/54) white. The DUB and PMP control groups consisted of 50% (6/12) black and 50% (6/12) white, respectively. Histopathological confirmation (62/66--94%) revealed 47/66 as negative, 5/66 as endometrial hyperplasia and 10/66 as squamous cell carcinoma or adenocarcinoma. A significant lesion with endometrial pathology was found in 23% of the patients. These findings suggest that the majority of women in this study with clinical symptoms of postmenopausal bleeding were negative for malignancies. While these results lean more towards a normal cytologic evaluation, postmenopausal bleeding should not be taken lightly. Postmenopausal bleeding could represent signs of more serious lesion such as squamous cell carcinoma or endometrial adenocarcinoma if not detected and managed early.

TCPL delivery devices: endometrial changes associated with exogenous sustained release of ovarian hormones.

The specific objective of this study was to investigate the ultrastructural changes in the female reproductive system exposed to long-term sustained release of medroxyprogesterone acetate (MPA), and MPA plus estrogen (MPA + E). Emphases were given towards the vaginal, endometrial and ovarian changes. The microcrystals of tricalcium-phosphate lysine (TCPL) were prepared by following standard laboratory procedure. Adult female rats (BW 250-300 gm) were randomly divided into three groups (n = 3). Group I animals were implanted with TCPL loaded with MPA (50 mg) plus E (50 mg), and group II animals were implanted with TCPL loaded with MPA (200 mg). Group III animals were left as our intact and unimplanted controls. At the end of 21 days post-implantation, all animals were sacrificed by means of an overdose of Halothane and the reproductive, as well as, the vital organs were collected, weighed, fixed, embedded, sectioned, and stained (H & E) for histopathological evaluations. Vaginal smears were collected prior the implantation to assess the fertility events and subsequently taken on a daily basis for the entire period of the investigation. The data obtained from this study demonstrates the following: (1) TCPL delivery system was capable of releasing MPA, and MPA + E at a sustained level for 21 days, (2) the use of TCPL devices resulted in the cessation of the estrus cycle and that was achieved at the end of 24 hour post-implantation, (3) remarkable physiological changes were observed in animals implanted with MPA, and MPA + E compared to control group animals, (4) regression of the ovarian tissue was observed in all MPA treated animals compared to intact control animals, and (5) histopathological evaluation of the endometrium of animals treated with sustained delivery of MPA, and MPA + E revealed increased stratification, altered polarity, cytoplasmic vacuolation and papillary projections.

The effects of dehydroepiandrosterone and dehydroepiandrosterone sulfate on the reproductive and vital organs of male rats.

Recent studies have documented that conventional administration (orally or injections) of DHEA (Dehydroepiandrosterone) or DHEAS (Dehydroepiandrosterone Sulfate) have induced alteration in tissues of the reproductive track of male rats. However, the exact mechanism of this physiological response has not been extensively studied. In addition, the route of DHEA or DHEAS administration has not been fully investigated. The specific objectives of this study were: (1) to deliver DHEA and DHEAS at a sustained level by means of TCPL delivery system, and (2) to evaluate the ultrastructural changes associated with sustained delivery of DHEA and DHEAS at the reproductive and vital organs level. A total of 12 adult male rats (BW 250-270 gm) were randomly divided into four equal groups. Groups 1-3 were implanted with TCPL ceramic capsules loaded with 200 mg DHEA (low dose), 600 mg DHEA (high dose) and 200 mg DHEAS, respectively. Aseptic surgical techniques were performed throughout the experiment. Blood (2 mls) was collected every other day for biochemical analysis. The weights were recorded bi-weekly. At the end of 21 days post-implantation, all control and experimental animals were sacrificed following standard lab procedure (overdose of Halothane). The reproductive and vital organs were collected, weighed, fixed, embedded, sectioned, and stained (H&E) for histological evaluations. Data obtained from this investigation suggest the following: (1) body weights between the experimental (DHEA and DHEAS) and control were unchanged, (2) weights of the reproductive were significantly different than controls, (3) kidney weights wee the only vital organ that was statistically different than controls, (4) prostatic tissue of the experimental group showed signs of atrophy, and (5) focal atrophy was also evident in the seminferous tubules and testes of DHEA and DHEAS treated rats. Overall conclusion of this study suggest that in male rats, the use of DHEA and DHEAS in a sustained delivery system seems to show some physiological changes in the vital and reproductive organs.

Histopathological evaluation of female reproductive tract exposed to sustained delivery of DHEA, and DHEA + E.

Dehydroepiandrosterone (DHEA) is a hormone produced by the adrenals that serves as a precursor for numerous steroid hormones. Conventional modes of DHEA administration were limited to injections and oral routes, which result in several drawbacks. This mandates the desire for the development of a different route of DHEA administration. Sustained delivery of DHEA by bioceramic capsules has not been fully explored. The objectives of this study were: (1) to deliver DHEA, and DHEA + estrogen in a sustained manner using tricalcium phosphate-lysine (TCPL) bioceramic capsules, and (2) to evaluate the morphological changes of reproductive and vital organs using female rats as a model. A total of twelve adult female rats (220-250 g) were randomly divided into four equal groups. Rats in Groups 1 and 2 were implanted with TCPL capsules containing 200 mg DHEA, and 600 mg DHEA (DHEA-HD), respectively. Group 3 animals were implanted with one TCPL capsule containing 200 mg of DHEA and a second TCPL capsule containing 50 mg estrogen, (DHEA + E). Group 4 represented the control group. Aseptic surgical techniques were utilized during i.p. implantation of the capsules. After implantation, body weights were recorded and blood (2 ml) samples were taken biweekly for 21 days. Pap smears were taken daily. At the end of 21 days, the animals were sacrificed using an overdose of halothane. The vital and reproductive organs were harvested, processed, embedded, sectioned and screened for cellular changes. Data obtained from these procedures revealed slight hypertrophy of the heart and kidneys. In DHEA + E implanted rats, a statistically significant increase (P < 0.05) was observed in the weights of the tubules, cervix, and uterine tissues compared to the control animals. Data obtained from this study demonstrates that the proliferative effect of sustained delivery of DHEA on the reproductive organs (ovary, cervix, uterus, and tubes) of female rats. This study provides more insights regarding the physiological alteration induced by sustained delivery of DHEA.

Cytomorphology of tyrosine-rich crystalloids in fine needle aspirates of salivary gland adenomas.

OBJECTIVE: To evaluate the presence of tyrosine-rich crystalloids (TRC) in fine needle aspiration (FNA) specimens of pleomorphic adenomas of salivary gland. STUDY DESIGN: FNA specimens from 12 patients were reviewed, and the percentage of cases showing TRC was established. The staining properties of the TRC were evaluated as well as spontaneous fluorescence under ultraviolet (UV) light. RESULTS: Of the 12 pleomorphic adenomas, 4 showed TRC (30%) in the smears. Among the eight cytologically negative cases there were two that showed a few TRCs on histology. All positive cases were from African American patients. TRC stained weakly with Papanicolaou stain. TRC were deep blue with Diff-Quik. They fluoresced under UV light. CONCLUSION: TRC could be detected in FNA specimens. They were best seen under UV light. The Papanicolaou technique stained TRC very pale, making them difficult to see. Diff-Quik stained TRC dark blue, mimicking deposits of dye. The amount of TRC in histology paralleled the detection rate in cytology.