Recovery and Enhancement of Faded Cleared and Double Stained Specimens

Cleared and stained specimens may become faded and/or opaque after long periods in preservative solutions. We developed a technique to revitalize faded and/or destained specimens regardless of their age. By adapting and revising Wassersug's procedure for differential staining of bone and cartilage, we successfully cleared and restained numerous small amphibian specimens of various ages. After rehydrating, specimens were bleached in potassium hydroxide and hydrogen peroxide, stained with alcian blue, and macerated with trypsin as needed. Alizarin red stain was applied or reapplied, although staining occasionally was unsuccessful in old formalin fixed specimens. After restaining, the specimens were dehydrated and placed in glycerol.     

Termite resistance of solid wood and plywood treated with quaternary ammonia compounds and common fire retardants

The ability of termites to attack solid wood and plywood treated with quaternary ammonia compounds and common fire retardants was evaluated. The plywood and solid-wood specimens treated with either monoammonium phosphate (MAP), diammonium phosphate (DAP), ammonium sulfate (AS), didecyl dimethyl ammonium chloride (DDAC), or didecyl dimethyl ammonium tetrafluoroborate (DBF) were subjected to termite resistance tests using the subterranean termites Coptotermes formosanus Shiraki under laboratory conditions. The lowest mass losses and the highest termite mortalities were obtained for the solid-wood and plywood specimens treated with DDAC and DBF. Higher termite mortalities were seen in the plywood specimens treated with the fire retardants when compared to the solid-wood specimens. The MAP, DAP, and AS treatments lowered the mass losses in both solid-wood and plywood specimens in comparison with control specimens; however, DBF and DDAC protected specimens well against termite attack at both concentration levels tested.     

Recovery and Enhancement of Faded Cleared and Double Stained Specimens

Cleared and stained specimens may become faded and/or opaque after long periods in preservative solutions. We developed a technique to revitalize faded and/or destained specimens regardless of their age. By adapting and revising Wassersug's procedure for differential staining of bone and cartilage, we successfully cleared and restained numerous small amphibian specimens of various ages. After rehydrating, specimens were bleached in potassium hydroxide and hydrogen peroxide, stained with alcian blue, and macerated with trypsin as needed. Alizarin red stain was applied or reapplied, although staining occasionally was unsuccessful in old formalin fixed specimens. After restaining, the specimens were dehydrated and placed in glycerol.     

Relining of prosthesis with auto‐polymerizing hard denture reline resins: effect of post‐polymerization treatment on flexural strength

Background: It has been suggested that microwave irradiation and prosthesis immersion in hot water after its polymerization may improve mechanical and viscoelastic properties of acrylic resins. Purpose: This study was proposed to verify the influence of microwave post‐polymerization (PP) treatment over the flexural strength of thermo‐polymerizing acrylic resin specimens (QC‐20) relined or not with two different composition hard chairside auto‐polymerizing reliners [Kooliner (K) and New Truliner (NT)]. Materials and Methods: For this study, 50 specimens of 64 × 10 × 3.3 mm were polymerized and distributed into five groups. G1 (control) specimens without relining and PP; G2 specimens relined with K, without PP; G3 specimens relined with NT, without PP; G4 specimens relined with K, with PP (microwave irradiation with 650 W for 5 min); G5 specimens relined with NT, with PP. Tests were performed on a universal testing machine Instron 4411 with compression speed of 5 mm/min. Results: Specimens of K without PP did not show statistically different results (p < 0.05) when compared with control. However, when submitted to PP these specimens showed a significant increase in flexural strength. Specimens of NT showed the lowest flexural strength of all groups, with or without PP when compared with control and K groups. Conclusion: Microwave PP (650 W for 5 min) proved to be an effective method of improving the flexural strength of K relined prosthesis. However, it did not seem to affect NT specimens.     

Application of the membrane filter technique to bromodeoxyuridine immunochemistry for exfoliative cytology

The membrane filter technique for smear specimens of tumors in bromodeoxyuridine (BrdU) immunochemistry is described. The staining results of Raji cells processed using the filter technique was compared with that obtained by the conventional cytospin method. Although the BrdU mean labeling index (LI) for in cytospin specimens was almost the same as the LI in membrane filter specimens, filter specimens showed excellent staining and less cell destruction compared with those processed by cytospin. Small amounts of tumor specimens such as squamous cell carcinoma and polymorphous low-grade adenocarcinoma also were processed using the membrane filter appliance. For squamous cell carcinoma, the LI for the filter specimens was 5.36 ± 0.38 and that of the paraffin sections was 5.56 ± 0.38. The membrane filter technique provided relatively undamaged specimens for exfoliative cytology and will be useful for immunohistochemical evaluation of tumor cells and for routine, noninvasive cytological screening.     

Detection of Candida antigen in bronchoalveolar lavage fluid

While bronchoalveolar lavage is frequently performed to evaluate immunocompromised hosts for infection, the significance of rare yeasts found on the cytologic examination of lavage fluid is unclear. This study used the latex agglutination method to test lavage fluids for Candida antigen to assess its usefulness in distinguishing Candida pneumonia from Candida colonization of the respiratory tract or oral contamination of the lavage specimen. Ninety-seven specimens from 87 patients were categorized on the basis of historical, microbiologic, cytologic and serologic data. Bronchoalveolar lavage fluids were positive for Candida antigen in 0 of 20 specimens from normal controls, 0 of 14 specimens from patient controls, 5 (36%) of 14 specimens from patients with Pneumocystis carinii pneumonia, 0 of 5 specimens from patients with gastrointestinal candidiasis, 0 of 9 specimens contaminated by oral-derived yeasts, 2 (10%) of 19 specimens from patients with probable Candida colonization and 15 (94%) of 16 specimens from patients with clinical and laboratory evidence of Candida pneumonia. We conclude that this test assists in the differentiation of Candida pneumonia from other situations in which yeasts are recovered by bronchoalveolar lavage.     

Angiogenesis in non-small cell lung cancer: microvessel area in needle biopsy in vascular tumor density

OBJECTIVE: To evaluate angiogenesis in non-small cell lung cancer (NSCLC) through immunohistochemistry with CD34 and computerized image analysis comparing the microvessel area in needle biopsies and surgical specimens. STUDY DESIGN: Core biopsies and surgical specimens from 28 patients with NSCLC were reviewed. Microvessels were highlighted by immunohistochemistry with anti-CD34. Tumor microvessel area was measured in digital photographs from hot spots of all samples. RESULTS: Average microvessel area among core samples was 5,093.6 microm2 (range, 233.4-17,916.8) and among surgical specimens was 3,599.3 microm2 (range, 376.9-9,514.0). There was strong correlation between overall microvascular area in biopsies and surgical specimens (r = 0.7; p = 0.0001). Mean area of core specimens was used to divide cases into groups of low and high vascular areas. A slightly stronger correlation was observed between biopsies and surgical specimens with low vascular areas (r = 0.84; p = 0.0001). CONCLUSION: There is strong correlation of microvessel area in core biopsies and respective surgical specimens in NSCLC. These data indicate that core specimens could be used to assess the extent of angiogenesis in NSCLC in the pretreatment phase.     

Differentially expressed glycoproteins in pre- and post-digital rectal examination urine samples for detecting aggressive prostate cancer

Urinary glycoproteins associated with aggressive prostate cancer (AG-PCa) were previously reported using post-digital rectal examination (DRE) urine specimens. To explore the potential of using pre-DRE urine specimens for detecting AG-PCa, we compared glycoproteins between pre- and post-DRE urine specimens, verified the previously identified post-DRE AG-PCa-associated urinary glycoproteins in pre-DRE urine specimens, and explored potential new glycoproteins for AG-PCa detection in pre-DRE urine specimens. Quantitative glycoproteomic data were acquired for 154 pre-DRE urine specimens from 41 patients with no cancer at biopsy, 48 patients with non-AG-PCa (Gleason score = 6), and 65 patients with AG-PCa (Gleason score 7 or above). Compared to glycopeptides from the post-DRE urine data, humoral immunity-related proteins were enriched in pre-DRE urine samples, whereas cell mediated immune response proteins were enriched in post-DRE urine samples. Analyses of AG-PCa-associated glycoproteins from pre-DRE urine revealed that the three urinary glycoproteins, prostate-specific antigen (PSA), prostatic acid phosphatase (ACPP), and CD97 antigen (CD97) that were previously identified in post-DRE urine samples, were also observed as AG-PCa associated glycoproteins in pre-DRE urine. In addition, we identified three new glycoproteins, fibrillin 1 (FBN1), vitronectin (VTN), and hemicentin 2 (HMCN2), to be potentially associated with AG-PCa in pre-DRE urine specimens. In summary, glycoprotein profiles differ between pre- and post-DRE urine specimens. The identified AG-PCa-associated glycoproteins may be further evaluated in large cohort of pre-DRE urine specimens for detecting clinically significant PCa.     

Cytologic findings in vitreous fluids. Analysis of 74 specimens

OBJECTIVE: To review the cytologic findings of vitreous specimens and propose a simplified approach to them. STUDY DESIGN: Seventy-four vitreous specimens from 60 patients obtained either during a pars plana vitrectomy or by vitreous aspiration were reviewed. Clinical correlation was obtained on all patients. RESULTS: Findings suggestive of a specific disorder were present in 30 specimens (41%); cytologic examination of the remaining 44 showed nonspecific changes. A lymphoproliferative disorder, the intraocular malignancy suspected most often in this series, was identified in eight specimens (11%). Large cell lymphomas were evident in 5 specimens, 2 specimens were suspicious for lymphoma, and 1 specimen was consistent with plasmacytoma. Twelve specimens (16%) contained hemorrhage. In rare instances, specific infectious agents, such as parasites (5%), bacteria (1%) and fungi (3%), could be identified. The diagnosis of viral infections required ancillary studies. Lens fragments were identified in four cases (5%), and a diagnosis of lens-induced endophthalmitis could be rendered in one case (1%). Changes consistent with sarcoidosis were present in 3% of cases. CONCLUSION: Based on this experience with vitreous specimens submitted for clinical reasons, we found that they could be divided into three broad diagnostic categories: inflammation/infection (54 specimens/41 patients), hemorrhage (12 specimens/12 patients) and malignancy (8 specimens/7 patients).     

The cytopathology of bone marrow transplantation

Bone marrow transplantation (BMT) is an increasingly effective treatment for patients with hematologic disorders and malignant neoplasms. From 1975 to 1986, 1,457 specimens were obtained for cytologic evaluation from 328 of the 635 patients who received BMTs at Memorial Sloan-Kettering Cancer Center. These specimens consisted of 1,049 cerebrospinal fluids (CSFs) from 265 patients, 292 bronchoscopy specimens from 92 symptomatic patients and 116 other specimens (including brushings from the liver and gastrointestinal tract, sputa, urines and cervico-vaginal smears). CSF specimens examined before and after BMT from 80 (30%) patients showed an increased number of benign, nonepithelial cells, which were mainly lymphocytic or histiocytic in origin. Malignant cells were detected in CSF specimens from 44 (17%) patients. Bronchoscopy specimens from 3 patients had suspicious cells present; those from 27 patients contained opportunistic organisms. Atypical epithelial or lymphoreticular cells were seen in bronchial specimens from 49 patients. All cytologic findings were correlated with the pertinent clinical information as well as biopsy and autopsy material, including histopathologic evidence of graft-versus-host disease. Cytologic evaluation, especially of bronchial and CSF specimens, was useful in diagnosing the presence of malignant neoplasms, infectious organisms, inflammatory responses, reactive lesions and cellular atypia due to treatment.     

Evaluation of possible decay and termite resistance of particleboard containing waste tire rubber

Particleboard specimens produced by adding waste tire rubber particles were assayed against white- and brown-rot fungi and termites in laboratory conditions. Particleboards were manufactured from a mixture of pine and poplar particles bonded with two different resins (melamine/urea formaldehyde [MUF] and polyisocyanate [PI]) by adding waste tire rubber particles at three different levels (10%/90%, 20%/80%, and 30%/70% by weight of waste tire rubber/wood). The particleboard specimens with waste tire rubber were not generally resistant against four fungi tested. Only MUF-containing specimens showed considerably better performance in decay resistance tests using the brown-rot fungus, Postia placenta; however, addition of waste tire rubber into those specimens did not provide resistance in comparison with control specimens without tire rubber. Formosan termites were also able to degrade particleboard specimens with waste tire rubber.     

Biological collections in an ever changing world: Herbaria as tools for biogeographical and environmental studies

Plant specimens stored in herbaria are being used as never before to document the impacts of global change on humans and nature. However, published statistics on the use of biological collections are rare, and ecologists lack quantitative data demonstrating the relevance to science of herbarium specimens. I found 382 studies with original data that used herbarium specimens to document biogeographical patterns or environmental changes. Most studies are less than 10 years old, and only 1.4% of the herbarium specimens worldwide have been used to answer biogeographical or environmental questions. The vast majority (82%) of papers dealt with vascular plants, but some studies also used bryophytes, lichens, seaweeds and fungi. The herbarium specimens were collected from all continents, but most of the studies used specimens from North America (40% of studies) or Europe (28%). Many types of researches (conservation, plant disease, plant invasion, pollution, etc.) can be conducted using herbarium specimens. Climate change, and especially phenological reconstructions, are clearly emerging research topics. By group, small herbaria (<100,000 specimens) are consulted as often as very large herbaria (>1,000,000 specimens) for biogeographical and environmental research, but in most cases, only large facilities provide specimens collected worldwide. The median number of specimens per study in papers using computerized collections (15,295) was much higher than for papers that did not include electronic data (226). The use of molecular analyses to investigate herbarium specimens is still relatively unexplored, at least from biogeographical and environmental points of view. Combined with recently developed procedures to correct biases, herbarium specimens might provide in the near future exciting additional spatio-temporal insights that are currently unimaginable.     

Demonstration of epithelium-associated microbes in the oesophagus of pigs, cattle, rats and deer

Scanning electron microscope or light microscope examination of oesphageal specimens demonstrated the association of microbial cells with the epithelial surface. Rod-shaped bacteria and cocci were seen in specimens obtained from pigs and rats, cocci in specimens from cattle, and rods     

Cytologic findings in homosexual males with acquired immunodeficiency

During a three-year period (1979 to 1982), 248 cytology specimens were obtained from 38 homosexual males who were admitted for treatment and had a variety of nonspecific symptoms, such as general malaise and weight loss. Twenty patients had Kaposi's sarcoma. Exfoliative and aspiration/brush specimens were prepared mainly from cerebrospinal fluids and tracheobronchial secretions. One hundred twenty-four specimens were of diagnostic value, exhibiting cytologic abnormalities: acute inflammation (11 specimens), marked cellular atypia (39 specimens), increased number of mature and immature lymphoreticular elements (31 specimens) and organisms such as parasites and fungi (43 specimens). Awareness of an epidemic of acquired immuno-deficiency-associated disease among male homosexuals, in conjunction with special processing and careful interpretation of cytologic specimens, contributed to the identification of organisms and the ruling out of malignancy. Despite prompt treatment, a swiftly progressive clinical course resulted in the death of 20 patients.     

Can Flow Cytometry Reduce the Workload For Cervical Screening? the Results of A Series of 622 Specimens

A simple method permitting the flow cytometric examination of cervical specimens has been developed and an assessment made of the feasibility of relying on this method to screen women for cervical neoplasia. Examination of four flow cytometric parameters showed differences between morphologically normal and abnormal specimens and allowed identification of a proportion of the normal specimens. The system had a false negative rate of 8%. Our experience with cervical specimens has revealed a number of problems associated with their examination by flow cytometry and these are discussed.     

The value of direct fluorescent antibody (DFA) testing for the detection of Pneumocystis Carinii In cytological specimens

A direct fluorescent antibody (DFA) method was compared with methenamine silver staining (MSS) for the detection of Pneumocystis carinii in 384 cytological specimens. DFA testing was more sensitive than the MSS, with P. carinii detected in 31 specimens with DFA and 24 with the MSS. Results of the two methods disagreed in 17 specimens, all of which were sputa. Twelve sputum specimens were DFA positive/MSS negative and five were MSS positive/DFA negative. It is concluded that the DFA technique, although relatively expensive, is simple to perform and offers superior sensitivity to the MSS. However, in sputum specimens the combined use of DFA and MSS leads to optimal sensitivity for the detection of P. carinii.     

Clinical Significance of Frizzled Homolog 3 Protein in Colorectal Cancer Patients

Frizzled homolog 3 receptor was up-regulated in several gastrointestinal cancers such as esophageal and gastric cancers. Moreover, frizzled homolog 3 has recently reported to be expressed in colorectal adenoma specimens. In the present study, we investigated the clinical significance of frizzled homolog 3 protein in colorectal cancer patients. Using immunocytochemical staining, frizzled homolog 3 expression was examined in 186 colorectal cancer specimens, 79 colorectal adenoma specimens, 133 colorectal polyp specimens, 127 colorectal cancer specimens with lymph node and/or distant metastasis, 310 specimens of various non-colorectal cancer metastatic carcinomas and 40 specimens with simultaneous occurrence of colorectal cancer, colorectal adenoma and colorectal polyp. Statistical analysis was used to correlate frizzled homolog 3 protein expression to the clinicohistopathological factors, recurrence/metastasis and survival after follow-up for 42 months in colorectal cancer patients. Frizzled homolog 3 protein was expressed in 100% colorectal cancer specimens, 89% colorectal adenoma specimens, 75% colorectal polyp specimens and 69% normal colorectal epithelial tissues. Moreover, frizzled homolog 3 immunocytochemical scores were highly correlated with colorectal cancer progression. Furthermore, frizzled homolog 3 was expressed in a comparatively lower percentage of metastatic hepatocellular carcinoma and metastatic renal clear cell carcinoma with focal and very weak staining than other metastatic tumor types. On the other hand, the frizzled homolog 3 immunocytochemical scores of colorectal adenomas with synchronous colorectal carcinomas were significantly higher than those of pure colorectal adenomas. Statistical analysis showed that frizzled homolog 3 immunocytochemical scores were associated with Dukes stage and lymph node status. Finally, stratified groups of colorectal cancer patients had significant differences in their recurrence/metastasis and survival. In conclusion, the present large-scale study has clearly showed that frizzled homolog 3 protein can generate clinically important information for colorectal cancer patients.     

细胞松弛素B对多头绒泡菌有丝分裂的影响

将细胞松弛素Ｂ（Ｃｙｔｏｃｈａｌａｓｉｎ Ｂ,ＣＢ）注入同步化的多头绒泡菌原质团,在光镜和电镜下跟踪观察有有丝分裂进程,发现多头绒泡菌进入有丝分裂的时间推迟,与未经ＣＢ处理的样品相比,在Ｓ期注入ＣＢ的样品进入有丝分裂的时间推迟３５ｍｉｎ,在Ｇ２早期注入ＣＢ的样品则推迟２０ｍｉｎ;在Ｇ２中期注入的推迟４５ｍｉｎ;在Ｇ２晚期注入的推迟６０ｍｉｎ,说明抑制肌动蛋白的功能则使有丝分裂受到明显影响。ＣＢ处理     

"Repair cells" in equine uterine cytologic and histologic specimens

Cells resembling those known as "repair cells" in gynecologic cytology specimens from women were identified in uterine cytology specimens from infertile mares treated with antibiotics using indwelling uterine catheters. This prompted a study of the effect on the equine uterus of indwelling catheterization without antibiotic infusion, using light microscopic examination of cytologic and biopsy specimens and electron microscopic examination of biopsy specimens. Cytologic and biopsy specimens had features within normal limits at the start of the study. Following five days of indwelling catheterization, neutrophils were present in both cytologic and biopsy specimens. In cytologic specimens, numerous groups of "repair cells" were present; similar cells in biopsy specimens indicated this was a focal reaction. The large nuclei and prominent nucleoli of the "repair cells" suggested cellular proliferation or regeneration. However, this was contradicted by the ultrastructural sparsity of ribosomes, endoplasmic reticulum, Golgi apparatus and mitochondria. Inflammation and "repair cells" were not present in cytologic or biopsy specimens collected 40 days after the start of the study. Although these cells may be a component of a repair process, our results support the hypothesis that "repair cells" in human and equine gynecologic cytology specimens are injured, rather than regenerating, cells. The term dysphaneroplastic (Greek: "abnormal cytosol development") is proposed to describe these cells since the cytoplasm does not reflect the features of cellular activity suggested by the nuclear appearance.