A pilot study of recovery room bypass ("fast-track protocol") in a community hospital.

PURPOSE: To evaluate the effectiveness of short-acting anesthetic drugs and techniques to achieve recovery room bypass criteria after minor surgery in a community hospital environment. METHODS: After agreement by a multidisciplinary committee, a pilot project was undertaken to assess the usefulness of ultra- short acting anesthetic drugs and pre-emptive analgesia to facilitate rapid recovery from general anesthesia. A cohort of 100 ASA I-II patients aged 18-65 yr undergoing simple knee arthroscopy or minor peripheral orthopedic procedures was compared to a similar cohort treated in the three months prior to the study period. Outcomes of interest included patient morbidity, success in achieving post-anesthesia care unit (PACU) bypass criteria, impact upon nursing resources, duration of operating room (OR) and hospital stay, and pharmaceutical costs before and after implementation. RESULTS: No patient morbidity was demonstrated prior to discharge home, and successful PACU bypass occurred in 83% of cases. Achievement of PACU discharge criteria while in the OR did not prolong the OR time, and discharge from hospital occurred earlier in the patients who did not require PACU care (P=0.0006 all "fast-track cases" vs all "controls"). Nursing complaints were more numerous when the day surgery personnel did not normally participate in PACU care. The cost of anesthetic care was significantly more using ultra-short acting drugs (CDN $14.17 vs CDN $20.57), but closer adherence to protocol could reduce this differential (CDN $18.84). CONCLUSION: Not all patients who receive a general anesthetic require admission to a phase I recovery facility. However, the justification for use of more expensive pharmaceuticals to achieve PACU bypass requires extensive changes in operating systems and voluntary professional behaviours.

Issues of quality and practicability in task analysis: preliminary results from two surveys.

Surveys of task analysis studies in two different industrial sectors within the UK, defence and nuclear power, have shown that there can be wide variability between them, both in terms of level of reporting and the depth of analysis. Particularly prevalent shortcomings were that data sources and methods were often either not specified or were described in insufficient detail. Thus users of the information could be asked to accept potentially costly design recommendations with little indication of the strength of the evidence, or of the effects of taking no action. Some tentative conclusions are drawn about the use of specific task analysis techniques but, while these are useful pointers, more evidence is required. Therefore, further study of task analysis techniques is needed in order to provide analysts with better guidance regarding the selection and use of task analysis methods and the reporting of task analysis findings.

Evaluation of culture systems containing bovine oviduct epithelial cells or granulosa cells to mature and maintain the developmental competence of bovine oocytes in vitro.

The effects of estrous cow serum (ECS), bovine oviduct epithelial cells (BOEC), and bovine granulosa cells (GC) on in vitro maturation (IVM) of immature oocyte-cumulus complexes (OCCs) were evaluated. Selected OCCs were cultured for 24 to 26 h in microdroplets of culture medium (CM; TCM 199 + 25 mM HEPES + 100 mug gentamicin sulfate/ml) or in CM medium supplemented or conditioned with 20% ECS, BOEC +/- 20% ECS or GC + 20% ECS. Supplemented media were incubated for 2 h before addition of OCCs, whereas media were conditioned by incubation with 20% ECS or BOEC +/- 20% ECS for 6 d, or with 20% ECS +/- GC for 24 or 48 h before addition of OCCs. The developmental competence of oocytes after TVM was assessed by insemination with glass wool separated, frozen-thawed bovine spermatozoa in microdroplets of modified medium (TALP) containing heparin (5 mug/ml) and BOEC for 18 h. The presumptive zygotes were cultured in microdroplets of CM medium + 20% ECS + BOEC for 7 to 9 d to assess embryo development to morula and blastocyst stages. The percentages of OCCs undergoing IVM (85 to 94%) and in vitro fertilization (IVF) (66 to 80%) were high, irrespective of the IVM conditions. Only after the IVM of OCCs in CM medium alone was the percentage of oocytes undergoing IVF significantly lower (66%; P<0.05). The proportion of IVF oocytes developing to blastocysts with a normal complement of cells (126 to 138) increased significantly (P<0.05) when the OCCs were matured in supplemented or conditioned CM medium containing ECS and/or somatic cells (18 to 28%) compared with those in CM medium alone (9%). When the CM medium was supplemented or conditioned with GC + 20% ECS, the proportion of fertilized oocytes developing to blastocysts increased significantly (28%; P<0.05). These results indicate that the potential of immature OCCs to be fertilized and to complete embryonic development to the blastocyst stage in vitro is enhanced by maturation in CM medium containing 20% ECS and/or BOEC or GC.

Developmental expression of the relaxin gene in the porcine corpus luteum.

Northern analysis and in-situ hybridization were used to follow the development of relaxin gene expression in the newly forming corpus luteum (CL) after ovulation and throughout luteal development. Alkaline phosphatase (AP) was used as a marker of theca-derived lutein cells and the relationship between AP-positive and relaxin mRNA-containing cells was assessed. Ovaries from prepubertal pigs treated with pregnant mares serum gonadotrophin (PMSG)/human chorionic gonadotrophin (hCG) were collected during the periovulatory period and at various times during 19 days after ovulation. In addition, CL from cyclic pigs on days 10 and 16 were used to monitor relaxin gene expression in small and large luteal cells. Northern analysis revealed that relaxin gene expression increased with CL development in the PMSG/hCG-treated pig, reaching maximal levels at around day 14 post-ovulation. Thereafter, as the CL regressed, the level of relaxin mRNA declined. In CL from cyclic pigs at day 10 of the cycle, only small luteal cells expressed relaxin mRNA. However, by day 16 of the cycle, large luteal cells were the source of relaxin gene expression. In-situ hybridization studies revealed that in the early CL (up to 30 h post-ovulation), the relaxin gene transcript was observed in cells along the margins of the CL and in the core of the infolding follicle wall corresponding to the AP-positive, luteinized theca cell layer. As luteinization progressed, the theca and granulosa cell layers could no longer be distinguished morphologically (from 54 h after ovulation until day 9). However, the pattern of relaxin hybridization persisted along the periphery in bands of cells penetrating the CL, and coincided with areas of AP staining, indicating that the theca lutein cells were the site of relaxin gene expression. At day 14, relaxin hybridization and AP staining were distributed throughout the luteal tissue. With CL regression both AP staining and relaxin hybridization declined. This pattern of relaxin hybridization in the CL of the gonadotrophin-primed pig was identical to that observed in cyclic pigs on days 10 and 16 of the cycle. These findings indicate that theca interna cells retain their ability to express the relaxin gene following ovulation and luteinization. In the early CL, the small theca-derived lutein cells are the source of relaxin transcript. However, as the CL becomes fully differentiated, the large granulosa-derived lutein cells acquire the capacity to express the relaxin message.

Sources and biological actions of relaxin in pigs.

Although the major source of relaxin in pigs is the corpus luteum of pregnancy, there is now evidence for relaxin gene expression and translation into protein in the theca interna cells of the preovulatory follicle, the corpus luteum of the cycle and the uterus. The theca interna cells retain their ability to express the relaxin gene and protein following ovulation. During the early stages of development of the corpus luteum, the theca-derived small lutein cells are the source of the relaxin transcript. As the corpus luteum becomes fully functional, there is a switch in the site of relaxin synthesis from small theca-derived lutein cells to large granulosa-derived cells. In the absence of luteolysis, this switch is accompanied by a dramatic rise in relaxin synthesis. Relaxin has been identified in boar seminal plasma and can maintain or increase sperm motility. However, a source of relaxin in the boar has not been identified. Relaxin is an important regulator of uterine function during pregnancy acting systemically to suppress myometrial activity and promote cervical dilation at parturition. The changes in thecal relaxin production during follicle development and its ability to promote growth and changes in proteolytic enzyme activity of granulosa cells in vitro have led to the concept of an autocrine or paracrine role for relaxin within the follicle. Uterotrophic effects of relaxin have been reported in rodents and swine and support the hypothesis that relaxin promotes uterine growth and expansion in early pregnancy to accommodate the growing fetuses. Mammotrophic effects of relaxin in rodents have now been extended to pigs, with evidence that relaxin is necessary for normal mammary parenchymal development in late pregnancy. In most instances the mechanisms responsible for, and the physiological significance of, these diverse biological effects remain to be elucidated.

Age at puberty, fertility and litter size of ewe lambs reared under different photoregimens.

Age at puberty, fertility and litter size of ewe lambs of synthetic sire and dam strains raised under different photoregimens were determined. The lambs were bred during January, May or September at 30 to 32 weeks of age. Irrespective of birth date, the lambs were reared under continuous light from birth to 5 weeks of age. From 5 to 20 weeks of age, they were kept under 16 hours of light dairy (16L:8D; Treatment A), 8 hours of light daily (8L:16D; Treatment B), or a split photoperiod of 8 hours total light daily (7L:9D:1L:7D; Treatment C). Subsequently, all lambs were exposed to 9 hours of light daily until after breeding. Lambs were exposed to rams for two estrous periods after treatment with fluorogestone acetate-impregnated intravaginal sponges and pregnant mares' serum gonadotropin (PMSG) to induce synchronized estrus. Although the age at puberty (174 days) was similar among treatments, the incidence of puberty prior to progestagen sponge treatment was higher (approximately 50%) for lambs reared under Treatments A and C than under Treatment B. Fertility and litter size of lambs were not influenced by the previous photoperiod history or by sexual maturity, i.e., puberal or prepuberal, at the start of the sponge treatment. However, strain, age and weight of lambs at breeding influenced significantly the reproductive outcome.

Relaxin gene expression in the porcine follicle during preovulatory development induced by gonadotrophins.

Northern analysis was used to identify relaxin gene expression in ovaries of prepubertal pigs primed with pregnant mare's serum gonadotrophin (PMSG) and human chorionic gonadotrophin (hCG). The cellular distribution of relaxin transcript in the developing follicle was localized by in-situ hybridization histochemistry. Three probes complementary to non-overlapping regions of the porcine prorelaxin molecule were used to identify relaxin gene expression in ovarian follicular tissue collected 0, 48, 60, 72 and 84 h after treatment with PMSG/hCG. A 1 kb transcript was detected in ovarian extracts of prepubertal gilts from 48 to 84 h after PMSG stimulation. This corresponds to the molecular size of the relaxin transcript reported in the pregnant sow ovary. Relaxin mRNA levels increased in ovaries from animals 48 through 84 h after PMSG. In-situ hybridization showed that the site of relaxin synthesis was the theca interna layer of the developing follicle. Relaxin mRNA was not observed in other follicular cell types, in small or atretic follicles or in follicles from unstimulated animals. The distribution and relative concentration of relaxin mRNA showed a good correlation with in-vitro production and immunohistochemical localization of relaxin previously reported in the developing pig follicle. The presence of both protein and mRNA for relaxin in the growing follicle supports a role for relaxin as a local regulator of ovarian function.

Changes in and partial identification of the plasminogen activator and plasminogen activator inhibitor systems during ovarian follicular maturation in the pig.

Plasminogen, plasmin, and plasminogen activator (PA) activities and PA and PA inhibitor (PAI) contents were measured in granulosa (GC) and theca interna cell extracts and follicular fluid (FF) obtained from preovulatory follicles of prepubertal gilts treated with eCG and hCG to induce follicular growth and ovulation. Plasmin activity in FF increased just before the time of expected ovulation. This increase was not attributable to changes in plasminogen levels, which remained relatively constant during preovulatory follicular development. The increase in follicular plasmin levels was associated with significant (p less than 0.01) increases in PA activity and content and decreases in PAI content in GC and FF. Western blot analysis suggested that follicular PA activity was represented principally by two forms of tissue type PA (t-PA) each with a pI of 7.8 and with molecular masses of 72,000 and 78,000 daltons, respectively. Two PA-PAI complexes of 126,000 and 130,000 daltons were observed. These complexes were partially dissociated with nucleophilic agents into two t-PA-like forms and a 52,000-dalton PAI protein with a pI of 4.8. Biochemical characteristics of the PAI protein suggest that it belongs to the same class of inhibitors as bovine and human PAI-1. These data indicate that rupture of the porcine ovarian follicle is temporally associated with a net increase in PA activity and an increase in plasmin activity. The increase in PA activity appears to be regulated by changes in PA and PAI content.

The synthesis and actions of steroids and prostaglandins during follicular maturation in the pig.

Our understanding of the synthesis and production of follicular steroids and prostaglandins (PG) in the pig is based largely on in-vitro studies with granulosa and theca interna tissues obtained from Graafian follicles at various stages of maturation. As the follicle enlarges before the LH surge, granulosa cells exhibit a decrease in FSH receptors and are less responsive to FSH in terms of cAMP production. Concurrently, there is an increase in granulosa and thecal cell LH receptors associated with an increase in responsiveness to LH and an increase in steroid production. Both granulosa and thecal cells produce oestrogen and progesterone, the rates of production being dependent on the stage of maturation of the follicle and substrate availability. Thecal cells are the principal source of androgens and control oestrogen synthesis by providing aromatizable substrate. After exposure to LH/hCG in vivo, both cell types lose the ability to produce oestrogen in vitro. These studies support the two-cell, two-gonadotrophin hypothesis of ovarian steroidogenesis. In vitro, granulosa and thecal cells exhibit an increased ability to produce PGE-2 and PGF-2 alpha after exposure to LH/hCG in vivo. Follicular PG production appears to be regulated by arachidonic acid availability and PG synthetase activity. In vivo, the follicular fluid concentrations of PGE-2 and PGF-2 alpha increase markedly at the time of ovulation. The increases in PG levels and ovulation can be blocked by indomethacin, an inhibitor of PG synthesis. These studies provide convincing evidence for an intrafollicular source of PGs and are consistent with the hypothesis that LH induces an increase in PG production that is essential for rupture of the follicle. Steroids act on the follicle through autocrine and paracrine mechanisms to modulate follicular growth and differentiation and to regulate steroidogenesis. PG actions on the follicle appear to be exerted via effects on contractile elements of the theca externa, blood vessels and on collagenolytic and other proteolytic enzymes.

Localization of relaxin during formation of the porcine corpus luteum.

The presence and localization of relaxin (RLX) in luteal tissue during the estrous cycle of the pig have been studied using the avidin-biotin immunoperoxidase method and homologous antisera to purified RLX. Prepubertal gilts were induced to ovulate by treatment with pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Ovaries were obtained at laparotomy during the periovulatory period and at specified times through Day 19 post-ovulation. Emphasis was placed on obtaining ovarian tissue at 12- and 24-h intervals up to 96 h after ovulation. RLX immunostaining was evident in theca interna (TI) cells before and at 6 h after ovulation. At 18 h after ovulation, RLX immunostaining comparable to that seen in TI cells was observed for the first time in luteinizing granulosa (G) cells. As luteinization progressed, it became difficult to identify the origin of the RLX immunostaining cells. However, the intensity of RLX immunostaining increased with corpus luteum (CL) development, with the staining becoming localized in the large luteal cells. By Day 19 after ovulation, RLX immunostaining was undetectable. These results indicate RLX is present in the CL during its formation and functional lifespan. Also, it would appear that the presence of RLX in G cells post-ovulation is associated with cell luteinization.

Gonadotropic regulation of prostaglandin production by ovarian follicular cells of the pig.

Prepubertal gilts were treated with 750 IU pregnant mare's serum gonadotropin (PMSG) and 72 h later with 500 IU human chorionic gonadotropin (hCG) to induce follicular growth and ovulation. Dispersed granulosa cells (GC) and theca interna cells (TC) from follicles of gilts 72 h (GC-72 and TC-72, respectively) and 108 h (GC-108 and TC-108 h, respectively) after PMSG treatment were cultured for 0, 12, 24, and 36 h in medium with or without luteinizing hormone (LH), dibutyryl cyclic adenosine 3',5'-monophosphate [Bu)2cAMP), calcium ionophore (A23187), and/or arachidonic acid (AA), and the production of prostaglandin E2 (PGE) and prostaglandin F2 alpha (PGF) was measured by radioimmunoassay. TC-72 was the principal source of PGs 72 h after PMSG. At 108 h, the production of PGE and PGF by GC was increased 10- and 30-fold, respectively, whereas corresponding increases by TC were 2-fold. LH and A23187 significantly stimulated PGE and PGF production by both GC-72 and TC-72, but only thecal PG production was stimulated by (Bu)2cAMP. LH had minimal or no effect on PG production by GC-108 and TC-108, but A23187 (GC-108, TC-108) and (Bu)2cAMP (TC-108) were stimulatory. Basal PG production by GC-72, GC-108, and TC-108 was stimulated by AA. However, production by GC and TC cultured in medium containing AA and LH, A23187, or (Bu)2cAMP was not different from that produced by AA alone. These findings suggested that GC and TC can synthesize PGs in vitro, but AA availability is rate-limiting in GC. After exposure to hCG in vivo, the capacity of both cell types to produce PGs is increased but is limited by AA availability.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of a new diluent and different processing procedures for cryopreservation of ram semen.

Ram semen was processed for freezing after initial dilution with a modified Tris-fructose diluent. Two aliquots were processed by cooling gradually to 5 degrees C, further dilution, equilibration and freezing in 0.5 ml straws either in pressurized liquid nitrogen (LN(2)) vapor (Method A) or on a block of dry ice (Method B). A third aliquot was cooled rapidly to 16 degrees C and then slowly to 5 degrees C, diluted further, equilibrated and frozen in straws in pressurized LN(2) vapor (Method C). The second dilution was carried out using a new diluent based on dextran-lactose. The diluted semen was equilibrated for 2 h before freezing. Semen was evaluated by artificial insemination (AI). The fertility of ewes bred by a double insemination with frozen-thawed semen processed by Methods A, B and C was 73% (n = 33), 67% (n = 30) and 80% (n = 30), respectively. In comparison, the fertility of ewes inseminated with fresh semen was 93% (n = 31). These preliminary data indicate an acceptable fertility can be achieved by AI with frozen-thawed semen processed using improved procedures.

Photoperiod entrainment of testosterone, luteinizing hormone, follicle-stimulating hormone, and prolactin cycles in rams in relation to testis size and semen quality.

Adult rams were exposed to photoperiod treatments over 2 years to study the influence of light regimes on pituitary-testicular activity and semen quality. Initially, all rams (12 per group) were exposed to 3 months of long days (16L:8D). Group 1 was then exposed to a regime of continuous short days (8L:16D) and Groups 2, 3, and 4 were exposed to 4 months of short days alternated with 1, 2, or 4 months, respectively, of long days. Every 2 weeks, serum hormone levels and scrotal circumference were determined and semen quality was evaluated. Regular cycles in pituitary and testicular activities corresponding to the period of the lighting regime resulted in Groups 2, 3, and 4, but not in Group 1. In general, the change from long days to short days induced increases in follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone levels, scrotal size and sperm numbers and a decrease in prolactin. The reverse occurred after subsequent exposure to long days. After 4 months of long days, testicular regression was complete, but when long-day exposure was reduced, less regression occurred. With continuous exposure to short days, FSH and testosterone remained above basal levels, prolactin levels were depressed, scrotal size remained near the maximum, and elevated numbers of motile sperm were sustained.

Secretion of 17 alpha-hydroxyprogesterone, androstenedione, and estrogens by porcine granulosa and theca interna cells in culture.

The steroid secreting activities of dispersed granulosa and theca interna cells from preovulatory follicles of prepubertal gilts 72 h after pregnant mare's serum gonadotropin treatment (750 IU) were compared. The cells were cultured for 24 h with or without steroid substrate (10(-8) to 10(-5) M progesterone, 17 alpha-hydroxyprogesterone, or androstenedione), FSH (100 ng/mL), LH (100 ng/mL), and cyanoketone (0.25 microM, an inhibitor of 3 beta-hydroxysteroid dehydrogenase). Granulosa cells cultured alone secreted mainly progesterone. Theca interna cells secreted mainly 17 alpha-hydroxyprogesterone and androstenedione, with secretion being markedly enhanced by LH. In the presence of cyanoketone, which inhibited endogenous progesterone production, theca interna but not granulosa cells were able to convert exogenous progesterone to 17 alpha-hydroxyprogesterone and androstenedione, and exogenous 17 alpha-hydroxyprogesterone to androstenedione and estradiol-17 beta in high yield. The secretion of the latter steroids from exogenous substrates was unaffected by LH. Theca interna cells secreted more estradiol-17 beta than did granulosa cells in the absence of aromatizable substrate, but estradiol-17 beta secretion by the latter was markedly increased after the addition of androstenedione. These apparent differences in steroid secreting activity between the cell types suggest that the enzymes responsible for conversion of C21 to C19 steroids, i.e., 17 alpha-hydroxylase and C17,20-lyase, reside principally in the theca interna cells. However, aromatase activity appears to be much higher in granulosa cells.

Localization of relaxin in the pig follicle during preovulatory development.

The avidin-biotin immunoperoxidase method and antisera to purified porcine relaxin were used to localize relaxin in sections of follicles from pregnant mare's serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-primed pigs during preovulatory development. Prepubertal pigs were treated i.m. with PMSG (750 IU) and 72 h later with hCG (500 IU) to induce follicular development and ovulation. Follicles were collected from untreated gilts or from gilts 24, 48, 60, 72, 84, 96, or 108 h after PMSG treatment. Light immunostaining in the theca interna was observed early in follicular development, at 48 and 60 h post-PMSG. At 72 h post-PMSG, relaxin immunostaining in the theca interna of the preovulatory follicle was more intense. After hCG treatment, the intense thecal immunostaining persisted and was apparent 84 and 96 h after PMSG. At about 6 h prior to expected ovulation (108 h post-PMSG), there was thinning of the follicle wall and a reduction in relaxin immunostaining in the theca interna. Immunoactive relaxin was not detected in follicles from untreated gilts, follicles 24 h post-PMSG, small healthy or atretic follicles, or in granulosa cells, theca externa or ovarian stroma, at any of the time points studied. These studies support the hypothesis that the theca interna is the primary source of follicular relaxin and provide further evidence for a paracrine role for relaxin in the ovulatory process.

Effect of PMSG dosage on the reproductive performance of adult ewes and ewe lambs bred at a progestagen-PMSG synchronized estrus.

The effect of dose of pregnant mares' serum gonadotropin (PMSG) on the reproductive performance of adult ewes and ewe lambs and lamb survival at birth after treatment with fluorogestone acetate (FGA)-impregnated intravaginal sponges and PMSG (250 IU or 500 IU) to synchronize estrus was evaluated. Ewes were exposed to rams for breeding at the synchronized and subsequent estrous cycles. The flock, comprised of three synthetic strains and two control breeds, was maintained in a controlled environment and exposed to an artificial light regimen which alternated at 4-mo intervals from 16h of light daily to 9h of light daily. Trials were conducted during January, May and September at the end of a 9-h daylength cycle. Adult ewes were bred in May and 8 mo later in January. Ewe lambs were bred in September at 6.5 to 7.5 mo of age. The overall reproductive performance of the adult ewes was similar at the two breedings: fertility approximately 90%, prolificacy approximately 2.7, fecundity approximately 240% and lambs born alive approximately 2.4. Dosage of PMSG had no effect. Reproductive performance of ewe lambs was lower and there was a strain x treatment interaction, suggesting greater variability in response. The results indicate there is no advantage to using a higher dose of PMSG in ewes with a natural relatively high fecundity. Moreover, the use of the artificial photoperiod appears to overcome the natural seasonal variation in reproductive performance.

Differential production of steroids by dispersed granulosa and theca interna cells from developing preovulatory follicles of pigs.

Dispersed granulosa and theca interna cells were recovered from follicles of prepubertal gilts at 36, 72 and 108 h after treatment with 750 i.u. PMSG, followed 72 h later with 500 i.u. hCG to stimulate follicular growth and ovulation. In the absence of aromatizable substrate, theca interna cells produced substantially more oestrogen than did granulosa cells. Oestrogen production was increased markedly in the presence of androstenedione and testosterone in granulosa cells but only to a limited extent in theca interna cells. The ability of both cellular compartments to produce oestrogen increased up to 72 h with androstenedione being the preferred substrate. Oestrogen production by the two cell types incubated together was greater than the sum produced when incubated alone. Theca interna cells were the principal source of androgen, predominantly androstenedione. Thecal androgen production increased with follicular development and was enhanced by addition of pregnenolone or by LH 36 and 72 h after PMSG treatment. The ability of granulosa and thecal cells to produce progesterone increased with follicular development and addition of pregnenolone. After exposure of developing follicles to hCG in vivo, both cell types lost their ability to produce oestrogen. Thecal cells continued to produce androgen and progesterone but no longer responded to LH in vitro. These studies indicate that several functional changes in the steroidogenic abilities of the granulosa and theca interna compartments occur during follicular maturation.