We report on the formulation and use of an alcoholic-agar additive solution generally useful for rapid, inexpensive liquid-based cytology slide preparation. Gynecological cytology specimens were collected from 1,000 women. Two hundred fifty aliquots each of CytoRich, CytoRich Red (Tripath Imaging, Inc., Burlington, NC), Preservcyt (Cytyc Corp., Boxborough, MA), and DINA*TRANS (DINA*CYT Corp., Portland, OR) fixatives were used for this study. Fixed cell suspensions from 1,000 women, seen consecutively by a general obstetrics and gynecology practice, were vortex-mixed and transferred into a premeasured amount of alcoholic-agar in test tubes. Test tubes were conventionally centrifuged, cells were trapped in a spontaneously formed agar-gel, and the supernatant solutions were decanted. Vortex-mixing cell buttons caused a rapid gel-to-sol transition, affording viscous cell suspensions that were applied to slides, smeared, and stained using Papanicolaou stains. Slides showed unclumped, monolayered, uniform, random cell-spreads. All of the fixatives afforded crisp presentation of normal and abnormal cells. There was an about 3-fold increase in HSIL+ (0.7-1.8%) and LSIL diagnoses (1.3-4.4%), and a 45% reduction in ASCUS diagnoses (3.3-1.8%), as compared to our cytology laboratory's previous year's patients' statistics with a concurrent 0.2% unsatisfactory rate, due solely to inadequate sampling. This manual method makes liquid-based cytology inexpensive and does not require specialized preparative devices.

The hormone relaxin has been implicated in the regulation of several processes in the reproductive tract during pregnancy and parturition. This study investigated the uterine effects of relaxin in immature and mature ovariectomized, estrogen-primed rats using morphometric and histochemical analysis. Rats were sprayed at 30 or 70 days of age and given estrogen (5 micrograms) 7 days later. After a week, they received an injection of porcine relaxin (100 micrograms) and were killed 6 h later; controls received vehicle alone. Histological sections were obtained from 7 levels of each uterine horn, and the volumes of endometrium and myometrium were calculated by use of a Zeiss Videoplan Computer Image Analyzer. In immature animals, relaxin treatment doubled uterine weights during the treatment period, and cross sections from relaxin-treated animals exhibited significant increases in the areas of both the myometrium and endometrium, 150% and 130% respectively. Mature animals were less responsive to relaxin although they also exhibited significant increases in uterine weight (31%), myometrial volume (29%), and endometrial volume (22%). With the use of Masson's Trichrome stain for collagen, we observed that relaxin alters the connective tissue framework of both endometrium and myometrium; control uterine collagen appears highly organized and dense with compact collagen fibers, whereas the collagen of relaxin-treated uteri is loosely arranged and disorganized with widely separated collagen fibers. Relaxin-stimulated uteri exhibited significantly greater vascularization, as evidenced by the size of arteries and veins in the vascular region between the circular and longitudinal muscle layers. Increased vascularization and uterine blood flow may be one mechanism involved in relaxin's uterotropic effect and is being investigated further.(ABSTRACT TRUNCATED AT 250 WORDS)

We report a technical improvement upon a previously disclosed manual liquid-based cytology (MLBC) method; and, we use the improved method to prepare slides from residual ThinPrep specimens in order to see how often ThinPrep diagnoses correspond to diagnoses derived from exhaustive examination of their parent sample suspensions. Residual cell suspensions from 500 ThinPrep cases comprising (1) 20 low-grade squamous intraepithelial lesions (LSILs); (2) 200 high risk (HR) negatives and 20 ASC-US; and (3) 260 screening cytology specimens were studied. Institutional review committee guidelines allowed us to know diagnoses by groups of specimens, but did not allow us to know individual patient diagnoses, so we could not perform case-by-case matched outcome-comparisons. Cells were concentrated by conventional centrifugation and sedimented into a polymer gel that was then vortex-mixed and converted into a viscous cell-rich suspension. The cell suspension was smeared between two clean glass slides, which were air-dried and stained with the Papanicolaou stain. Two study-sets were created, comprising one slide from each case. Each of the two study sets was examined by two cytopathologists, and discordant diagnoses were adjudicated. Because of the ambiguity involved in the "atypical" (ASC-US, ASC-H, AGC) diagnosis categories, only outcomes at the level of LSIL or greater were recorded. All MLBC SILs were digitally imaged and abnormal slides plus digital images were sent to the laboratory that provided the residual automated liquid-based cytology (ALBC) suspensions. The final diagnoses were confirmed by the laboratory that provided the residual ALBC specimens. MLBC slides of the 20 LSIL cases afforded 2 high-grade squamous intraepithelial lesions (HSILs) and 18 LSILs. Those of the 200 HR-Negatives showed 3 HSILs and 30 LSILs; and those of the 20 HR-ASC-US showed 3 HSILs and 9 LSILs. MLBC slides of the 260 screening cytology specimens showed 1 Carcinoma, 3 HSILs and 20 LSILs; affording 3 HSILs and 14 LSILs more than originally diagnosed. The MLBC method of this report is useful for preparing cell suspensions for cytological examination. Our analytical method was exhaustive and used nearly all of the cell material that was provided to us for analysis; therefore, we conclude that this approach is useful for determining how well ALBC instruments represent their parent sample suspensions. It appears that "rare events" may be overlooked when limited sample aliquots are analyzed by ALBC instruments, and this probably accounts for our increased discovery of SILs by the MLBC method. Also, SILs often present as aggregates of cohesive cells which, if overlooked or ineffectively transferred to ALBC slides, would not be diagnosed.

Previously spermatozoa in the semen of vasectomized men were reported in 62 of 63 specimens from 24 men 2 to 31 years postvasectomy (Freund and Couture, 1982). A morphologic basis and term, "microrecanalization," was proposed for this observation. Serial sections (5 mu at 200-mu intervals) of 40 specimens removed at vasovasostomy from 20 men (2 to 14 years postvasectomy) were examined and microcanals (small epithelial-lined channels) were demonstrated in 27 specimens from 18 men. In nine of the 27 specimens, spermatozoa or sperm heads were found within the microcanals. Microcanals occurred in smooth muscle, connective tissue and scar tissue, in each segment, testicular, central and abdominal, in the presence or absence of the vas deferens. Microcanal continuity was traced for 200 to 1140 microns by computerized image analysis. Microrecanalization is characterized by the absence of inflammation or sperm extravasation and is histologically distinct from vasitis nodes or sperm granuloma. Microrecanalization provides morphologic and physiologic bases for the protection of the testis and maintenance of spermatogenesis in man after vasectomy.

We show that residual cell material from ThinPrep PapTest (Cytyc Corporation, Boxborough, MA) atypical squamous-cells of undetermined significance (ASCUS) cases may be manually reprocessed to triage women into actionable diagnostic categories (HSIL, LSIL, and Negative). Material remaining from each of 358 ThinPrep ASCUS cases was manually reprocessed as two slides, labeled "A" and "B." Interobserver agreement between case contributors (CCs) and three sequential reviewers (SRs) was analyzed with 186 cases (Study 1), and diagnostic reproducibility between SRs was examined with an additional 172 cases (Study 2). In Study 1, CCs classified 33% of cases as LSIL or greater, SRs classified 60% as LSIL or greater, and there was 58% diagnostic agreement between CCs and SRs. No "Negative" case assignment by one group afforded an "HSIL" assignment by the complementary group. In Study 2, there was 95% agreement between SRs A slide and B slide diagnoses with 54% of A slides and 55% of B slides classified as LISL or greater. Again, no "Negative" case assignment to one slide afforded an "HSIL" assignment to the complementary slide. Overall, 12.6% of the 358 cases showed HSIL, and all HSILs by one observer group were ASCUS or greater by the complementary observer group. Using manual reprocessing beyond the 21-day specimen outdate for HPV testing by the Hybrid Capture II High Risk HPV test (HR-HCII; Digene Corporation, Beltsville, MD), many ThinPrep ASCUS cases were reclassified as LSIL or HSIL. The 12.6% HSIL proportion of this study approximated the 11.5% CIN 2 or greater proportion of the ALTS ASCUS arm. Similar to ALTS, manual liquid-based cytology (MLBC) would have referred nearly 60% of women to colposcopy for a definitive diagnosis of HSIL or LSIL without resorting to HPV testing. These data demonstrate that many cases of automated liquid-based cytology (ALBC)-diagnosed ASCUS represent unrecognized SIL, which is present in the ALBC specimen vial at the time the ASCUS diagnosis is rendered.