MANAGEMENT OF EARLY PROSTATIC CARCINOMA

It is important to make a diagnosis of prostatic carcinoma as early as possible, because early treatment gives the best results whether radical prostatectomy is done or endocrine therapy used. Open perineal biopsy is the most accurate method of making a diagnosis. Perineal needle biopsy or the newer approach of transrectal needle biopsy is probably about 75 per cent accurate in making a diagnosis.Ten-year survival with conservative therapy, as determined in a review of a series of cases, was 50 per cent—about the same as that following radical prostatectomy; but the patients with prostatectomy are clinically free of malignant disease whereas the former are not. Radical prostatectomy is indicated in a few selected cases.The results from endocrine therapy begun immediately after diagnosis are significantly better than those from delayed treatment. Orchiectomy and estrogens promise a little longer survival than estrogens alone.     

Sentinel node detection and radioguided occult lesion localization in breast cancer

Sentinel lymph node biopsy might replace complete axillary dissection for staging of the axilla in clinically N0 breast cancer patients and represent a significant advantage as a minimally invasive procedure, considering that about 70% of patients are found to be free from metastatic disease, yet axillary node dissection can lead to significant morbidity. In our Institute, Radioguided Occult Lesion Localization is the standard method to locate non-palpable breast lesions and the gamma probes is very effective in assisting intra-operative localization and removal, as in sentinel node biopsy. The rapid spread of sentinel lymph node biopsy has led to its use in clinical settings previously considered contraindications to sentinel lymph node biopsy.In this contest, we evaluated in a large group of patients possible factors affecting sentinel node detection and the reliability of sentinel lymph node biopsy carried out after large excisional breast biopsy. Our data confirm that a previous breast surgery does not prohibit efficient sentinel lymph node localization and sentinel lymph node biopsy can correctly stage the axialla in these patients.     

Fibroblast sources: Where can we get them?

Fibroblasts are cells widely used in cell culture, both for transient primary cell culture or permanent as transformed cell lines. Lately, fibroblasts become cell sources for use in disease modeling after cell reprogramming because it is easily accessible in the body. Fibroblasts in patients will maintain all genetic background during reprogramming into induced pluripotent stem cells. In spite of their large use, fibroblasts are obtained after an invasive procedure, a superficial punch skin biopsy, collected under patient’s local anesthesia. Taking into consideration the minimum patient’s discomfort during and after the biopsy procedure, as well as the aesthetics aspect, it is essential to reflect on the best site of the body for the biopsy procedure combined with the success of getting robust fibroblast cultures in the lab. For this purpose, we compared the efficiency of four biopsy sites of the body (skin from eyelid, back of the ear, abdominal cesarean scar and groin). Cell proliferation assays and viability after cryopreservation were measured. Our results revealed that scar tissue provided fibroblasts with higher proliferative rates. Also, fibroblasts from scar tissues presented a higher viability after the thawing process.     

A method of processing first-trimester chorionic villous biopsies for cytogenetic analysis

Chorionic villous biopsy is emerging as a technique for obtaining fetal cells for prenatal diagnosis in the first trimester of pregnancy. Chromosome analysis has been performed on small villous biopsies using either direct harvests of uncultured cells or after culturing villous tissue. Here, we describe a method where both techniques can be used simultaneously; from a single villous biopsy, GTG-banded chromosomes of improved morphology are obtained from direct preparations and from cultured villous cells.     

Swab or biopsy samples for bioburden testing of allograft musculoskeletal tissue?

Swab and biopsy samples of allograft musculoskeletal tissue are most commonly collected by tissue banks for bacterial and fungal bioburden testing. An in vitro study was performed using the National Committee for Clinical Laboratory Standards standard ‘Quality control of microbiological transport systems’ (2003) to validate and evaluate the recovery of six challenge organisms from swab and biopsy samples of allograft musculoskeletal tissue. On average, 8.4 to >100 and 7.2 to >100 % of the inoculum was recovered from swab and biopsy samples respectively. A retrospective review of donor episodes was also performed, consisting of paired swab and biopsy samples received in this laboratory during the period 2001–2012. Samples of allograft femoral heads were collected from living donors during hip operations. From the 3,859 donor episodes received, 21 paired swab and biopsy samples each recovered an isolate, 247 swab samples only and 79 biopsy samples only were culture positive. Low numbers of challenge organisms were recovered from inoculated swab and biopsy samples in the in vitro study and validated their use for bioburden testing of allograft musculoskeletal tissue. Skin commensals were the most common group of organisms isolated during a 12-year retrospective review of paired swab and biopsy samples from living donor allograft femoral heads. Paired swab and biopsy samples are a suitable representative sample of allograft musculoskeletal tissue for bioburden testing.     

Accuracy of the analysis of multiple small fragments of glial tumors obtained by stereotactic biopsy.

To examine the reliability of the diagnoses reached on multiple small fragments of cerebral glial tumors obtained via stereotactic biopsy, samples obtained from 100 consecutive glial tumors (during real or simulated biopsy) were studied by cytology and histology. In comparison to the definitive diagnosis made on the whole tumor, a correct positive diagnosis on the biopsy sample was made by histology in 96% of cases and by cytology in 93% of the cases (with 96% correct results when combining both methods). A correct identification of the tumor type and grade was achieved by histology in 82% of cases and by cytology in 80% of the cases (with 85% correct results when combining both methods). The limits of stereotactic biopsy are related to the difficulty of identifying all of the typical tumor features on tiny tissue fragments of a pleomorphic neoplasm, such as a glioma. This study demonstrates that better results may be obtained by using both cytology and histology to study multiple stereotactic biopsy samples from glial tumors.     

CARCINOMA IN SITU OF THE UTERINE CERVIX—DIAGNOSIS BY BIOPSY

In 50 cases of carcinoma in situ of the cervix uteri, the lesion was present in the cervical canal in 36 instances, but the squamous epithelium of the portio was involved in only 14 cases. Since single biopsy from the portio or external os may show no malignant change even in cases in which vaginal smears are positive for cancer, single biopsy is indicated only in the presence of gross suspicious lesions.When positive smears have been obtained, cold-knife cone biopsy is indicated (a) whenever gross lesions are not visible on the ectocervix, (b) if carcinoma in situ is found in a biopsy of the external os, in order to determine the presence or absence of invasion, and (c) when there are repeated positive smears and biopsy of the portio has not shown the presence of malignancy.The technique of cone biopsy is given in detail.     

Ultrasound-guided diagnostic breast biopsy methodology: retrospective comparison of the 8-gauge vacuum-assisted biopsy approach versus the spring-loaded 14-gauge core biopsy approach

Background

Ultrasound-guided diagnostic breast biopsy technology represents the current standard of care for the evaluation of indeterminate and suspicious lesions seen on diagnostic breast ultrasound. Yet, there remains much debate as to which particular method of ultrasound-guided diagnostic breast biopsy provides the most accurate and optimal diagnostic information. The aim of the current study was to compare and contrast the 8-gauge vacuum-assisted biopsy approach and the spring-loaded 14-gauge core biopsy approach.

Methods

A retrospective analysis was done of all ultrasound-guided diagnostic breast biopsy procedures performed by either the 8-gauge vacuum-assisted biopsy approach or the spring-loaded 14-gauge core biopsy approach by a single surgeon from July 2001 through June 2009.

Results

Among 1443 ultrasound-guided diagnostic breast biopsy procedures performed, 724 (50.2%) were by the 8-gauge vacuum-assisted biopsy technique and 719 (49.8%) were by the spring-loaded 14-gauge core biopsy technique. The total number of false negative cases (i.e., benign findings instead of invasive breast carcinoma) was significantly greater (P = 0.008) in the spring-loaded 14-gauge core biopsy group (8/681, 1.2%) as compared to in the 8-gauge vacuum-assisted biopsy group (0/652, 0%), with an overall false negative rate of 2.1% (8/386) for the spring-loaded 14-gauge core biopsy group as compared to 0% (0/148) for the 8-gauge vacuum-assisted biopsy group. Significantly more (P < 0.001) patients in the spring-loaded 14-gauge core biopsy group (81/719, 11.3%) than in the 8-gauge vacuum-assisted biopsy group (18/724, 2.5%) were recommended for further diagnostic surgical removal of additional tissue from the same anatomical site of the affected breast in an immediate fashion for indeterminate/inconclusive findings seen on the original ultrasound-guided diagnostic breast biopsy procedure. Significantly more (P < 0.001) patients in the spring-loaded 14-gauge core biopsy group (54/719, 7.5%) than in the 8-gauge vacuum-assisted biopsy group (9/724, 1.2%) personally requested further diagnostic surgical removal of additional tissue from the same anatomical site of the affected breast in an immediate fashion for a benign finding seen on the original ultrasound-guided diagnostic breast biopsy procedure.

Conclusions

In appropriately selected cases, the 8-gauge vacuum-assisted biopsy approach appears to be advantageous to the spring-loaded 14-gauge core biopsy approach for providing the most accurate and optimal diagnostic information.     

Diagnosis of lymphoma by image-guided needle biopsies: fine needle aspiration biopsy, core biopsy or both?

OBJECTIVE: To determine whether or not concurrent core biopsy adds to results obtained from image-guided fine needle aspiration biopsy (FNAB) in cases of lymphoma. STUDY DESIGN: Twenty-eight FNABs of lymphomas with adjuvant flow cytometry (FC) and concurrent core biopsy were evaluated retrospectively. In each case, completeness of diagnosis by FNAB, including phenotyping and grading, where appropriate, was reviewed. The contribution of core biopsy to the diagnosis in cases where FNAB did not render a complete diagnosis was assessed. Prognostic information not available from the FNAB but obtained from the core biopsy was also evaluated. RESULTS: FNAB with adjuvant FC gave a complete diagnosis, including phenotype and grade, where applicable, in 23 of 28 cases (82%). Core biopsy added to the diagnosis in 3 cases. In 1 case, large B-cell lymphoma was diagnosed on core biopsy when FNAB was unsatisfactory. In the other 2 cases, grade of follicle center cell lymphoma was higher on core biopsy than on FNAB. The addition of the information obtained by core biopsy to that obtained by FNAB raised the diagnostic accuracy to 93%. Core biopsy was used to assess nodularity, which could not be determined on FNAB. Core biopsy was also used to assess prognostic markers by immunohistochemistry (Ki-67 and p53); they were not available with FC. This was done in 11 cases when requested by the oncologist. CONCLUSION: FNAB with adjuvant FC is a useful technique for diagnosing and subtyping lymphomas. However, diagnosis and subclassification are often insufficient. Core biopsy material provides opportunity for obtaining additional diagnostic and prognostic information that may not be easily derived from the FNAB. This allows optimal treatment planning in patients for whom excisional biopsy is contraindicated.     

Small bowel biopsy for malabsorption: comparison of the diagnostic adequacy of endoscopic forceps and capsule biopsy specimens.

Biopsy specimens of the small bowel were obtained from 40 patients suspected of having malabsorption. Four different techniques were used at a single session--namely, endoscopic biopsy of the descending duodenum using paediatric and standard size forceps and suction capsule biopsy of the descending duodenum and the proximal jejunum. Specimens were compared for size, adequacy, and ability to confirm or exclude mucosal abnormality. Fourteen patients had villous atrophy. In all patients four biopsy specimens were obtained with paediatric endoscopic forceps and four with standard endoscopic forceps. No capsule biopsy specimen was retrieved from the duodenum in three patients and from the jejunum in five patients. Specimens were considered to be adequate in 36 patients when paediatric forceps were used, in 39 when standard forceps were used, in 28 on duodenal capsule biopsy, and in 32 on jejunal capsule biopsy. This study indicates that the most reliable method for diagnosing or excluding villous atrophy is endoscopic forceps biopsy of the descending duodenum, provided that at least four specimens are obtained with standard size forceps.     

Chemiluminescent measurement of telomere DNA content in biopsies

Telomeres are nucleoprotein complexes that protect the ends of chromosomes from fusion and degradation. They are typically shorter in tumor cells than in paired normal cells, and shorter telomeres are associated with poor outcome in cancer. We previously described a slot blot-based methodfor measuring telomere DNA content, a proxy for telomere length. Although this method represented an improvement over existing methods, its 30-ng limit of sensitivity was insufficient for use with biopsy or other scant tissues. Here we describe a chemiluminescent slot blot assay for telomere DNA content that has the sensitivity required for use with biopsy materials. The results obtained with DNA derived from human placental, HeLa, human peripheral blood lymphocytes, sham-needle core prostate biopsies, and archival prostatectomy tissues demonstrated that telomere DNA content can be reliably and reproducibly measured in 5 ng, and sometimes as little as 2 ng, genomic DNA. Sham-needle core prostate biopsy and prostatectomy specimens processed in parallel produced comparable results. The contribution of truncated telomeres in admixtures containing as much as 75% normal placental DNA could be established. We also demonstrated that the treatment of tissue with formalin before DNA purification does not decrease the efficacy of the assay.     

Detection of DNA in Human Blastocyst Cavity Aspirate by Multiplex PCR

Preimplantation genetic testing (PGT) is a modern method of detection of chromosomal and genetic abnormalities in a human embryo before its transfer to the uterus. The genetic material is obtained by embryo biopsy. Here, we attempted to evaluate the efficiency of a method of noninvasive biopsy—aspiration of the blastocoel contents (blastocentesis) of human embryos. In this study, a biopsy was carried out human embryos with low morphological characteristics (3–4 CC according to Gardner grading) on day 6–7 of development. DNA obtained from the aspirate, as well as from the blastocyst, was analyzed by QF-PCR (quantitative fluorescent polymerase chain reaction) after whole genome amplification. In total, 24 blastocysts and aspirate samples obtained from them were analyzed; assayable DNA was found in seven (29%) aspirate samples from the blastocyst cavity, and this DNA was identical to blastocyst DNA in five (71%) cases. Thus, it was shown that, using aspiration of the blastocoel fluid of the human embryo, it is possible to obtain DNA suitable for analysis by molecular genetic methods. The features and advantages of the use of multiplex QF-PCR method combined with whole genome amplification for studying DNA obtained during aspiration of the blastocoel fluid are discussed. The prospects of DNA obtainment by the noninvasive biopsy method for preimplantation genetic testing (PGT) in the routine practice of infertility treatment and prevention of chromosomal and genetic abnormalities in newborns are considered.     

ORAL MUCOUS MEMBRANE LESIONS—Pathologic Features

Lesions of the oral mucous membranes often present bizarre, clinical and histological patterns because of the salivary environment and the complex organization of the various membranes. Diagnosis of oral lesions depends primarily on tissue biopsy, which must be modified in some cases from the usual dermatological technique. Exfoliative cytology has become an important adjunct to biopsy. Congenital, keratotic, malignant and dermatological lesions are those which are most easily diagnosed by clinical as well as histopathological and cytological examination.The first professional examination of an oral lesion is by far the most important because at this time the determination of the correct diagnostic procedure is made.     

Needle biopsy of the liver. A critique of four currently available methods.

There are currently four needle biopsy methods for obtaining tissue from patients with possible diffuse liver disease or cancer. These include percutaneous blind needle biopsy, a visually guided needle biopsy at laparoscopy, guided fine-needle biopsies with ultrasonography or computed tomography, and the transvenous liver biopsy. We and others have found the guided fine-needle biopsy technique to be safe, relatively cheap, and highly accurate in the diagnosis of liver cancer. Blind percutaneous biopsy should be reserved for patients with possible diffuse, noncancerous, liver disease. Guided biopsies at laparoscopy can be done if the other two methods fail to give a tissue diagnosis. The transvenous approach is useful in patients with a coagulation disorder.     

Liquid biopsy for early diagnosis of non-small cell lung carcinoma: recent research and detection technologies

Liquid biopsy, an innovative method for early diagnosis of cancer, has changed the traditional method of diagnosing lung cancer and is considered a feasible auxiliary diagnostic tool. To date, various reports emphasize the need for non-small cell lung carcinoma (NSCLC), both with higher incidence and mortality and less effective treatments; thus, emphasizing the need for early detection of NSCLC for improved patient outcomes. Invasive tissue biopsy is a common diagnostic tool that is usually extracted from the primary tumor to indicate molecular composition. In comparison, liquid biopsy taken from body fluid reflects extensive malignant features nonexistent in primary tumors. Owing to new detection technologies, liquid biopsy reduces the need for invasive treatments and enhances the accuracy and specificity of early detection of cancer in clinical settings. This review summarizes some latest research on the diagnosis of early-stage NSCLC via liquid biopsy, including circulating DNA, circulating tumor cells, exosomes, and tumor-educated platelets, as well as their detection technologies, such as fluorescence in-situ hybridization-based, polymerase chain reaction-based, next-generation sequencing-based, Chip-based, and microfluidic methods. Additionally, we outline the existing challenges and possible solutions for liquid-biopsy biomarkers. Our study mainly highlights the merits of liquid biopsy as a promising biomarker for non-invasive detection in the future, particularly for the early detection of NSCLC, thereby benefitting human health.     

"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.     

Overview of the role of liquid biopsy in cancer management

With the emergence of novel targeted therapeutic options in early-stage and advanced-stage malignancies, researchers have shifted their focus on developing personalized treatment plans through molecular profiling. Circulating tumor DNA (ctDNA) is a cell-free DNA (ctDNA) fragment, originating from tumor cells, and circulating in the bloodstream as well as biological fluids. Over the past decade, many techniques were developed for liquid biopsies through next-generation sequencing. This alternative non-invasive biopsy offers several advantages in various types of tumors over traditional tissue biopsy. The process of liquid biopsy is considered minimally invasive and therefore easily repeatable when needed, providing a more dynamic analysis of the tumor cells. Moreover, it has an advantage in patients with tumors that are not candidates for tissue sampling. Besides, it offers a deeper understanding of tumor burden as well as treatment response, thereby enhancing the detection of minimal residual disease and therapeutic guidance for personalized medicine. Despite its many advantages, ctDNA and liquid biopsy do have some limitations.This paper discusses the basis of ctDNA and the current data available on the subject, as well as its clinical utility. We also reflect on the limitations of using ctDNA in addition to its future perspectives in clinical oncology and precision medicine.     

DNA distribution in biopsy specimens from human cervical carcinoma investigated by flow cytometry.

Flow cytometry was used for the investigation of the DNA distribution in biopsy specimens from 51 patients with cervical carcinoma. Portio biopsy specimens from 9 pregnant women and from 10 patients with cancer of the breast served as controls. The results demonstrate that most specimens from patients suffering from cervical carcinoma contain considerable cell populations with increased DNA as compared with controls. The possible clinical significance of these findings is discussed.     

Prostate biopsy: current status and limitations

The technique of prostate biopsy has evolved over the past 10 years to improve our ability to detect prostate cancer. Extended biopsy schemes can be performed in the office under local anesthesia and are well tolerated. In addition to detection, the role of extended biopsy schemes in refining tumor grading and risk assessment has become better defined. This review discusses the evolution of prostate biopsy techniques from the sextant scheme to the extended scheme and demonstrates the latter's utility in clinical decision making.     

Endoscopic brushing cytology and biopsy in the diagnosis of upper gastrointestinal tract lesions. A study of 350 cases

Direct-vision endoscopic examination conducted on 4,000 patients for persistent upper gastrointestinal (GI) complaints over a period of five years revealed 350 visible lesions that were subjected to brushing cytology and biopsy. Cytologic examination of brushing smears from all 350 cases showed malignant cells in 67 (19.14%), cells suggesting benign polypoid neoplasms in 4 (1.14%), ulcerative and reparative features with attendant atypias in 186 (53.14%), inflammatory findings in 91 (26%) and false-negative findings in 2 cases (0.57%). Only 259 (74%) of the visible lesions were also subjected to endoscopic biopsy. Of the 67 patients with positive cytology, 52 were judged positive on the biopsy specimen; the 2 false-negative cytologic reports were confirmed as positive by biopsy. In four patients with gastric ulcers, malignant cells were seen along with gastric repair cells. This study indicates that brushing cytology is very useful in detecting benign ulcerative lesions with their atypias, a feature that could be useful in monitoring and controlling lesions in high-risk groups of patients, such as in India. In this study, endoscopic brushing cytology gave a better diagnostic yield than did endoscopic tissue biopsy. However, the two techniques are complementary for the diagnosis of upper GI malignancies.