Testicular fine needle aspiration cytology for the diagnosis of azoospermia and oligospermia

OBJECTIVE: To evaluate qualitative and quantitative cytologic features on testicular fine needle aspiration biopsy in the diagnosis of azoospermia and oligospermia and to correlate cytologic and histologic diagnoses. STUDY DESIGN: In this prospective study, 50 infertile males selected from the infertility clinic of Guru Tegh Bahadur Hospital were studied. Fine needle aspiration cytology (FNAC) smears from both testes of 27 azoospermic and 23 oligospermic patients (sperm count < 10 million per milliliter) were stained with May-Grünwald-Giemsa and Papanicolaou stain. Differential counting of 500 spermatogenic cells was done, and the number of Sertoli cells per 500 germ cells was determined for calculating the spermatic index and Sertoli cell index, respectively. FNAC and testicular biopsy were performed under local anesthesia as a minor surgical procedure. RESULTS: Six groups were identified on FNAC smears from azoospermic patients: I. normal spermatogenesis (8), II. hypospermatogenesis (2), III. maturation arrest (2), IV. Sertoli cells only (6), V. atrophic pattern (7), and VI. Leydig cell predominance (2). In oligospermic patients two groups were identified: I. those with normal spermatogenesis (4), and II. those with subnormal spermatogenesis (19). Correlation with histopathologic examination was seen in 81.5% azoospermic and 65.2% oligospermic patients. CONCLUSION: Qualitative and quantitative evaluation of testicular FNAC provides useful information on both azoospermic and oligospermic patients. FNAC performed under local anesthesia is an acceptable outpatient procedure that consistently yields sufficient diagnostic material in all patients.     

Correlation of cell counts and indices in testicular FNAC with histology in male infertility.

OBJECTIVE: To evaluate the value of percentage cell counts and cell indices in testicular fine needle aspiration cytology (FNAC) in male infertility and their correlation with histologic categories as seen in open testicular biopsies. STUDY DESIGN: Differential cell counts were performed, and cell indices, including spermatic index, Sertoli cell index and sperm-Sertoli cell index, were calculated in testicular fine needle aspiration (FNA) smears in 30 azoospermic males whose open testicular biopsies were classified as normal spermatogenesis in 10 cases, maturation arrest in 5, hypospermatogenesis in 6, Sertoli cell only syndrome in 5 and tubular/peritubular sclerosis in 4. RESULTS: In normal spermatogenesis, FNA smears showed up to 40% Sertoli cells, and spermatozoa were the predominant spermatogenetic cell type. There was a progressive increase in Sertoli cell percentage and Sertoli cell index and reduction in spermatozon percentage, spermatic index and sperm-Sertoli cell index with increasing severity of reduction in spermatogenesis in different histologic categories. The differences between mean counts and indices in normal spermatogenesis and other histologic categories were statistically significant (P < .01). CONCLUSION: The percent cell counts and cell indices in testicular FNAC correlate with histologic categories and are useful in evaluating male infertility.     

Touch imprint cytology in biopsy of the infertile testis.

OBJECTIVE: To identify different cell types in the testis by using touch imprint cytology and to compare the cytologic findings to the histopathologic diagnosis in infertile men. STUDY DESIGN: This prospective study used touch imprint preparations and included 20 infertile men. The biopsy material obtained was stained with toluidine blue, May-Grünwald-Giemsa stain and Papanicolaou stain. The cytologic results for oligospermic, normospermic and azospermic men were compared to the specific histopathologic diagnosis. The proportion of spermatogenic versus Sertoli cells was calculated. The scores were compared between three groups based on the results of the histologic biopsy: normal spermatogenesis, hypospermatogenesis and incomplete spermatogenic arrest. RESULTS: The mean ratio of the spermatogenic cells versus Sertoli cells was statistically significantly different in the three groups (P < .01). The mean ratio of spermatogenic cells to Sertoli cells was higher in cases with normal spermatogenesis than in cases with hypospermatogenesis and incomplete spermatogenic arrest, revealing a statistical difference (P<.01). This ratio was not statistically significantly different between the hypospermatogenesis and incomplete spermatogenic arrest groups. CONCLUSION: A cytologic demonstration of germinal cells by using touch imprint preparations may be a guide for histologic diagnosis.     

Les molécules de signalisation dans la physiologie et la pathologie testiculaire

Infertility affects an estimated 10% of couples, and in roughly half of these cases the defect can be traced to the men. Male infertility can be due to a failure of spermatogenesis (non-obstructive azoospermia) or to an obstruction (obstructive azoospermia) on deferent ducts. Non-obstructive azoospermia affect about 2% of men, they are due to a failure in spermatogenic maturation. The cause of these defects are still unclear, however, differents causes have been pointed out: genetic defect (Klinefelter syndrome, chromosome Y deletion), physical (irradiation), chemical or infectious affections. But in 40 to 60% of the cases, the cause of non-obstructive azoospermia is not determined. During the last years, from fundamental research, it was hypothezised that an alteration in extracellular signaling systems could potentially caused male infertility. Genetic manipulations show that knock-out or overexpression of some gonadic factors affect spermatogenesis (spermatogenic arrest or tumor formation), for example the systems TGFβ and SCF/c-kit. Experimental data obtained fromin vitro andin vivo approaches show that intratesticular signaling molecules regulate gonadic fonctions with hormones (LH/testosterone, FSH). These local factors might regulate testicular development in the fetal period with the genes of development. Moreover, through adulthood, these local factors could regulate spermatogenesis as a relay of the hormonal action in the testis. Finally, recent studies suggest that abnormalities in local factor expression could lead to testicular pathologies. Futur studies would certainly confirm the important role of signaling molecules in human testicular pathology.     

Localization of follicle-stimulating hormone (FSH) immunoreactivity and hormone receptor mRNA in testicular tissue of infertile men

Testicular biopsies from 82 oligo-or azoospermic male patients were subjected to immunostaining using anti-human FSH antibodies. Histological evaluation showed normal spermatogenesis (nspg) in 7 (FSH: 2.7±0.7), mixed atrophy (ma) in 63 (FSH:5.3±0.5), and bilateral or unilateral Sertoli Cell Only syndrome (SCO) in 12 (FSH:21.7±3.5) patients. For the relationship between FSH values and testicular histology, see Bergmann et al. (1994). FSH immunoreactivity was found exclusively in Sertoli cells and in some interstitial cells. Seminiferous epithelium showing normal or impaired spermatogenesis displayed only weak immunoreactivity compared to intense immunoreaction, i.e. large and numerous vesicles in Sertoli cells of SCO tubules in biopsies showing mixed atrophy or SCO. In addition, h-FSH receptor mRNA was demonstrated by in situ hydridization using biotinylated cDNA antisense oligonucleotides. Hybridization signals were found within the seminiferous epithelium exclusively in Sertoli cell cytoplasm associated with normal spermatogenesis and in epithelia showing different signs of impairment, including SCO. It is concluded that: (1) Sertoli cells are the only cells within the seminiferous epithelium expressing FSH receptors; (2) the accumulation of FSH immunoreactivity in Sertoli cells of SCO tubules appears to be a sign of impaired Sertoli cell function.     

Fine needle aspiration cytology in the management of male breast masses. Nineteen years of experience.

OBJECTIVE: To determine the utility and accuracy of fine needle aspiration cytology (FNAC) as well as its sensitivity, specificity and predictive value in the diagnosis of male breast masses. STUDY DESIGN: Data on male breast FNAC done between 1978 and 1997 were retrieved from the records of the cytopathology laboratory. FNAC diagnoses were categorized as positive, negative, inconclusive or unsatisfactory. Cytohistologic correlation was done with data from histopathology records. Sensitivity, specificity, diagnostic accuracy and predictive values of FNAC were calculated using standard statistical methods. RESULTS: Five hundred seven of 13,175 patients undergoing breast FNAC were males. Of them, 393/507 had satisfactory aspirates. Of these, 70 were positive (13.8%), 295 were negative (58%), and 29 were inconclusive (5.7%). A total of 114 FNACs (22.5%) were unsatisfactory. Histopathology was available in 97/507 cases. There were no false positive or false negative diagnoses. FNAC had a sensitivity, specificity and diagnostic accuracy of 100% for male breast lesions. CONCLUSION: This large study shows that FNAC is a very accurate tool for diagnosis of male breast lesions. It is highly sensitive and specific, with good cytohistologic correlation. FNAC should therefore be an integral part of the primary assessment of breast lumps in males.     

Utility of fine needle aspiration cytology in mediastinal lesions: a clinicopathologic study of 1617 cases from a single institution

OBJECTIVE: To study the distribution, prevalence and cytomorphologic features of mediastinal lesions and assess the utility of fine needle aspiration cytology (FNAC) in such lesions by correlating with clinical, laboratory and imageologic parameters. STUDY DESIGN: A retrospective study was performed of mediastinal lesions that were referred for ultrasonographic/computed tomography-guided FNAC during the years 2001-2006. Correlation was done wherever possible using the following parameters: histology, bone marrow, imageology, tumor markers, cytology, immunohistochemistry, antecedent history and regression after therapy. RESULTS: A total of 161 patients underwent 182 aspirates. Diagnosis was possible in 130 (80.7%) patients, and, in 31 cases (19.3%), aspirates were unsatisfactory. In 71 (54.6%) correlation was done, and in 70 (98.5%) positive correlation was found. CONCLUSION: FNAC in correlation with clinical, imageologic and hematologic features proved to be an excellent diagnostic tool in diagnosing as well as classifying mediastinal lesions and can be used as a substitute to core biopsy.     

Immunolocalization of inhibin and activin alpha and betaB subunits and expression of corresponding messenger RNAs in the human adult testis

Inhibin B is a testicular peptide hormone that regulates FSH secretion in a negative feedback loop. Inhibin B is a dimer of an alpha and a beta(B) subunit. In adult testes, the cellular site of production is still controversial, and it was hypothesized that germ cells contribute to inhibin B production. To determine which cell types in the testes may produce inhibin B, the immunohistochemical localization of the two subunits of inhibin B were examined in adult testicular biopsies with normal spermatogenesis, spermatogenic arrest, or Sertoli cell only (SCO) tubules. Moreover, using in situ hybridization with mRNA probes, the mRNA expression patterns of inhibin alpha and inhibin/activin beta(B) subunits have been investigated. In all testes, Sertoli cells and Leydig cells showed positive immunostaining for inhibin alpha subunit and expressed inhibin alpha subunit mRNA. Using inhibin beta(B) subunit immunoserum on testes with normal spermatogenesis and with spermatogenic arrest, intense labeling was located in germ cells from pachytene spermatocytes to round spermatids but not in Sertoli cells. Inhibin beta(B) subunit mRNA expression was intense in germ cells from spermatogonia to round spermatids and in Sertoli cells in these testes. In testes with SCO, high inhibin beta(B) subunit mRNA labeling density was observed in both Sertoli cells and Leydig cells, whereas beta(B) subunit immunostaining was negative for Sertoli cells and faintly positive for Leydig cells. These results agree with the recent opinion that inhibin B in adult men is possibly a joint product of Sertoli cells and germ cells.     

Fine needle aspiration cytology of the testis as the first-line diagnostic modality in azoospermia: a comparative study of cytology and histology

Objectives:  Male factors are responsible for about half of all infertility cases. Until recently, testicular biopsy was the standard method to ascertain the aetiology of azoospermia. Fine needle aspiration cytology has gained increasing popularity as a simple and minimally invasive procedure that can help in assessing testicular function accurately. This study was aimed at addressing the question whether testicular fine needle aspiration (FNA) may be used as a first-line diagnostic modality in azoospermia and to assess its usefulness in the diagnostic protocol.
Methods:  The FNA was performed in 78 consecutive azoospermic patients. To obviate sampling errors both testes were aspirated, except when contraindicated. Routine haematoxylin and eosin as well as Romanowsky staining was performed on the smears.
Results:  The smears were categorized on cytological examination into normal spermatogenesis in 35 (50%) patients, Sertoli cells only syndrome in 22 (31.4%) and maturation arrest at the spermatocyte/spermatid level was seen in 13 (18.4%) patients. There were eight (10.2%) cases with scant smears where cytological diagnosis could not be made. A good correlation between cytological smears and histological sections was found in 54 of 58 testes (93.1%) in which histopathological confirmation was available.
Conclusions:  Testicular FNA may be utilized as a first-line investigative modality in patients with azoospermia, provided the procedure is performed and interpreted by experts.     

Diagnostic accuracy of fine needle aspiration cytology of the thyroid: impact of ultrasonography and ultrasonographically guided aspiration

OBJECTIVE: To determine the impact of ultrasonography on the diagnostic accuracy of fine needle aspiration cytology (FNAC) of the thyroid. STUDY DESIGN: We compared two FNAC series. In period 1 we used neither ultrasonograph nor ultrasonographically-guided FNAC in the evaluation of thyroid nodules, while both tools were routinely applied in period 2. Moreover, in the former period all cellular follicular lesions were encountered among suspicious findings, while in period 2 we advised regular follow-up examinations instead of immediate surgery for patients with no significant atypia. The basis of follow-up examinations was ultrasonographic volumetry of the nodule and repeat FNAC on growing nodules. RESULTS: The number of FNAC, positive predictive value of FNAC, number of malignancies, sensitivity, the specificity and diagnostic accuracy were higher in period 2 as compared with period 1: 3,446 vs. 1,448; 45% vs. 16%, 39 vs. 29, 92% vs. 76%, 65% vs. 87% and 66% vs. 87%, respectively. CONCLUSION: The introduction of ultrasonography permits higher diagnostic accuracy of the evaluation process. This is achieved not only by lessening false negative reports but, more important, by decreasing false positive results.     

Fine needle aspiration of epididymal nodules in Chandigarh, north India: an audit of 228 cases

BACKGROUND: Epididymal nodules are not infrequently encountered in surgical practice. These are generally small and slippery and fine needle aspiration cytology (FNAC) is not easy. But as it is rapid and less traumatic than a biopsy, this is a favoured technique in the assessment of epididymal nodules. PATIENTS AND METHODS: In the present study, all the cases of epididymal nodules aspirated from January 1998 to August 2004 were retrieved from the cytology files of the Department of Cytology, Post Graduate Institute of Medical Education and Research, Chandigarh, India. RESULTS: A total of 228 cases were retrieved and divided as follows: tuberculous epididymitis 70 (30.7%), non-specific inflammation 10 (4.4%), microfilaria 2 (0.9%), hydrocele 26 (11.4%), spermatocele 42 (18.4%), spermatic granulomas 12 (5.3%), adenomatoid tumour 3 (1.3%), leiomyosarcoma 1 (0.4%) and lipoma 1 (0.4%). Thirty-six (15.8%) cases were labelled as benign aspirate not otherwise specified. FNAC material was inadequate for opinion in 22 (9.65%) cases and three (1.3%) cases revealed evidence of a haematoma. CONCLUSIONS: FNAC was useful in the diagnosis of 90.3% of cases, thereby avoiding surgical biopsy and other investigations. Therefore, FNAC has an important role in the differential diagnosis of epididymal nodules as it can detect malignancy and benign conditions such as tuberculosis and acute and chronic epididymo-orchitis.     

Cytopathologic diagnosis of xanthogranulomatous cholecystitis and coexistent lesions. A prospective study of 31 cases

OBJECTIVE: To evaluate the diagnostic accuracy and reliability of preoperative ultrasound (US)-guided fine needle aspiration cytology (FNAC) in the diagnosis of xanthogranulomatous cholecystitis (XGC) and coexistent lesions (carcinoma) and also to evaluate the possibility ofmissing either carcinoma or XGC on cytology. STUDY DESIGN: The cytologic diagnoses of XGC and coexistent lesions were made according to standard criteria. In a prospective, 5-year study, preoperative US-guided FNAC from 42 cases of XGC was compared with follow-up histologic diagnoses, which were available in 31 cases. When FNAC after the first aspiration showed the aspirate to be nondiagnostic, FNAC was repeated under US guidance. RESULTS: Preoperative US-guided FNAC diagnoses of XGC were made in 31 cases, for which follow-up histology was available in all cases. US-guided FNAC diagnosis ofXGC only was made in 30 cases and coexistent lesions in 1 case. Followup histology revealed 26 cases of XGC, 4 of a coexistent lesion and 1 of squamous cell carcinoma only. The overall diagnostic accuracy of preoperative US-guided FNAC was 96.77%. The overall possibility of missing XGC was 3.33% and that of carcinoma, 12.01%. CONCLUSION: Preoperative US-guided FNAC is safe, rapid, reliable, cost-effective and accurate in diagnosing XGC. However, the possibility ofcoexistent carcinoma cannot be definitely ruled out. It is therefore recommended that FNAC be performed from multiple suspicious sites under radiologic guidance. Thus, preoperative US-guided FNAC diagnosis would help in determining the urgency of treatment and also in planning the surgical procedure for gallbladder lesions.     

Pitfalls in the diagnosis of Wegener''s granulomatosis on fine needle aspiration cytology

OBJECTIVE: To review the clinical and pathological findings in six suspected cases of Wegener's granulomatosis (WG) and highlight the diagnostic difficulties faced by the cytopathologist. METHODS: Retrospective review of records of the Cytopathology Department to identify patients who underwent image-guided transthoracic pulmonary fine needle aspiration cytology (FNAC) for pulmonary lesions of suspected WG and those who were subsequently confirmed to have WG. Detailed evaluation of cytomorphological features was carried out. RESULTS: A total of six cases were identified in whom the initial procedure to obtain a pathological diagnosis was transthoracic FNAC. In one case, atypical squamous cells on cytology initially suggested a diagnosis of squamous cell carcinoma while in another a diagnosis of WG was made on cytology; however, a subsequent lung biopsy revealed silicosis. CONCLUSION: Acute inflammation and necrosis are the most consistent cytopathological findings in WG. In selected cases FNAC can provide supportive pathological evidence to establish a diagnosis of WG.     

Fine needle aspiration cytology of thyroid nodules: how accurate is it and what are the causes of discrepant cases?

Fine-needle aspiration cytology (FNAC) is widely accepted as the most accurate, sensitive, specific and cost-effective diagnostic procedure in the assessment of thyroid nodules and helps to select people preoperatively for surgery. The purpose of this study was to evaluate the results of thyroid FNAC in our institution and to determine the reasons for discrepancies between the cytological and histological diagnosis. We evaluated the cytological and histological results of 254 FNACs obtained from 231 patients who underwent subsequent thyroid surgery. All of the material was blindly reviewed for quality control, by one experienced cytopathologist. All FNACs were carried out under ultrasound guidance. The cytological diagnosis was classified as benign, suspicious, malignant or unsatisfactory. The definitive histological study showed benign lesions in 195 of the 231 patients (84%). A benign diagnosis based on FNAC was correct in 105 of the 108 benign cases (97%). FNACs diagnosed as 'suspicious' resulted in a distribution of 49 benign (79%) and 13 malignant (21%) diagnoses. FNAC showed malignancy in 34 cases (13%) and in only one case did the final histology differ from cytology (correlation 97%). The percentage of FNACs that were inadequate for diagnosis was 20%. Review of cytological and histological slides did not lead to any change in the original diagnosis. Our study revealed a cytological-histological discrepancy (2%) in only 4 out of 231 cases over a period of 10 years, due to either a diagnostic or sampling error.     

无精和严重少精症患者的遗传缺陷分析——-附2例世界首报染色体异常核型

对217例无精和严重少精症患者外周血淋巴细胞染色体核型进行分析,并采用聚合酶链反应对7例Y染色体结构异常患者的AZFc区进行检测。发现187例无精症患者中检出异常核型77例（41.18%）（其中46,XY,t(6;14)(p21;p13),46,XY,t(8;12)(p21;q24)为世界首报核型）,主要涉及染色体异常（数目异常和结构异常）;染色体异态（Y染色体异态和9号染色体臂间倒位）及46,XX性反转;30例严重少精症患者中检出异常核型4例（13.33%）（结构异常和46,XX性反转）。由此可见,性染色体数目和结构异常是精子发生障碍的主要原因,其次常染色体的某些断裂点也可能影响精子发生。AZFc区的缺失与否与精子发生也有直接关系。     

Expression of connexin 43 in human testis

In order to further characterize the Sertoli cell state of differentiation, we investigated the expression of connexin 43 (cx43) protein in the testis of adult men both with normal spermatogenesis and associated with spermatogenic impairment, since cx43 is first expressed during puberty. Cx43 protein was found as a single 43-kDa band on western blots of extracts of normal human testicular material. Cx43 immunoreactivity was generally present between Leydig cells. Within the normal seminiferous epithelium cx43 immunoreactivity was localized between adjacent Sertoli cells, except at stages II and III of the seminiferous epithelial cycle when primary spermatocytes cross from the basal to the adluminal compartment suggesting a stage-dependent Sertoli cell function. While testes with hypospermatogenesis and spermatogenic arrest at the level of round spermatids or spermatocytes revealed a staining pattern similar to that of normal adult testis, the seminiferous tubules showing spermatogenic arrest at the level of spermatogonia and Sertoli-cell-only syndrome were completely immunonegative. We therefore assume that severe spermatogenic impairment is associated with a population of Sertoli cells exhibiting a stage of differentiation deficiency. Accepted: 10 June 1999     

Fine needle aspiration cytology diagnosis, flow cytometry immunophenotyping and histology in clinically suspected lymphoproliferative disorder: a comparative study

OBJECTIVE: To evaluate advantages and drawbacks of fine needle aspiration cytology (FNAC) with flow cytometry (FC) in our routine, using, whenever possible, histology as the gold standard. STUDY DESIGN: From November 2003 to April 2005, we studied, by FNAC and FC, 113 patients with a tentative clinical diagnosis of lymphoproliferative disorder. Excision was performed in 43 patients. RESULTS: Excluding the 7 cases in which FNAC/FC made the diagnosis of metastatic carcinoma, a conclusive diagnosis was obtained with FNAC/FC in 87.7% (93 of 106) of patients. Most of these (n = 48) corresponded to reactive processes. Histologic study of 8 cases confirmed FNAC/FC diagnosis of reactive process. Insufficient material was obtained in 8 (7.1%) patients, and discordance between FNAC and FC occurred in 5 (4.4%), leading to inconclusive diagnosis. There was concordance in benign and malignant diagnoses between FNAC/FC and histology in every case in which conclusive diagnosis of FNAC/FC was advanced. CONCLUSION: FNAC and FC together provide a reliable, definitive diagnosis in most cases, obviating, whenever a reactive process is found, unnecessary surgery or follow-up. Histology was useful in the few cases in which FNAC/FC could not reach conclusive diagnosis and in subclassification of specific lymphomas.     

Pathology of supraclavicular lymphadenopathy in Chandigarh, north India: an audit of 200 cases diagnosed by needle aspiration

OBJECTIVE: Fine needle aspiration cytology (FNAC) of lymph nodes can be used routinely as a first-line diagnostic test. The majority of studies reveal a malignant cause for palpable supraclavicular lymph nodes. The present audit further emphasizes the use of FNAC as a first-line investigation for the evaluation of enlarged supraclavicular lymph nodes. METHODS: A total of 200 cases of palpable supraclavicular lymph node(s) were included in the present study. RESULTS: Left supraclavicular lymph nodes were found to be more commonly involved (59.5% cases). Sixty-four per cent cases showed metastatic deposits and 13.5% cases were diagnosed as tuberculosis. Ten per cent cases showed reactive lymphoid hyperplasia; 0.5% (one) case showed only necrosis and on autopsy, a microscopic focus of choriocarcinoma was found in the testis. In 7.5% cases, diagnostic material could not be aspirated despite repeated attempts. Common metastatic tumours were from lung (22% cases), breast (16.4% cases), cervix (11% cases) and oesophagus (8.6% cases). In 13.3% cases the primary site was unknown and the diagnosis of malignancy first came from FNAC. CONCLUSION: The present study further highlights the importance of FNAC as a first-line diagnostic modality in the evaluation of supraclavicular lymphadenopathy. A full history, radiological findings and immunochemistry in difficult cases can help to arrive at a definitive diagnosis.     

Fine needle aspiration cytology of lymph node: experience of 2 university hospitals with conventional smears and liquid-based cytology

OBJECTIVE: To compare 2 different series of fine needle aspirations (FNA) performed by conventional smears (CS) and liquid-based cytology (TriPath PREP). STUDY DESIGN: From January 1, 2001, to December 31, 2005, we selected 139 FNA samples of lymph node, if the diagnosis was histologically proven after the initial cytologic diagnosis. Samples came from 2 university hospitals from Brussels: UZ-Brussel (n=96) using liquid-based cytology (LBC) and Hospital Erasme ULB (n=43) using CS. RESULTS: The number of inadequate samples was greater for LBC than CS, but there was no statistical difference (17.7% vs. 4.7%, p = 0.059). No differences were found between LBC and CS in sensitivity (respectively, 85.0% vs. 85.2%), specificity (84.2% vs. 85.7%) and efficiency (84.8% vs. 85.4%). CONCLUSION: Despite the cost, the efficiency of lymph node FNA cytology is identical between the CS and LBC performed by the TriPath PREP system. The quality of the smears, both LBC and CS, depends mainly on practitioner dexterity. Nevertheless, one advantage of LBC is that it provides an ancillary technique to improve FNAC diagnosis. In the future, a combination of these technologies with FNAC alone could in some cases take the place of surgical lymph node excision.     

Defining regions of the Y-chromosome responsible for male infertility and identification of a fourth AZF region (AZFd) by Y-chromosome microdeletion detection

Cytogenetic and molecular deletion analyses of azoospermic and oligozoospermic males have suggested the existence of AZoospermia Factor(s) (AZF) residing in deletion intervals 5 and 6 of the human Y-chromosome and coinciding with three functional regions associated with spermatogenic failure. Nonpolymorphic microdeletions in AZF are associated with a broad spectrum of testicular phenotypes. Unfortunately, Sequence Tagged Sites (STSs) employed in screening protocols range broadly in number and mapsite and may be polymorphic. To thoroughly analyze the AZF region(s) and any correlations that may be drawn between genotype and phenotype, we describe the design of nine multiplex PCR reactions derived from analysis of 136 loci. Each multiplex contains 4-8 STS primer pairs, amplifying a total of 48 Y-linked STSs. Each multiplex consists of one positive control: either SMCX or MIC2. We screened four populations of males with these STSs. Population I consisted of 278 patients diagnosed as having significant male factor infertility: either azoospermia, severe oligozoospermia associated with hypogonadism and spermatogenic arrest or normal sperm counts associated with abnormal sperm morphology. Population II consisted of 200 unselected infertile patients. Population III consisted of 36 patients who had previously been shown to have aneuploidy, cytological deletions or translocations involving the Y-chromosome or normal karyotypes associated with severe phenotype abnormalities. Population IV consisted of 920 fertile (control) males. The deletion rates in populations I, II and III were 20.5%, 7% and 58.3%, respectively. A total of 92 patients with deletions were detected. The deletion rate in population IV was 0.87% involving 8 fertile individuals and 4 STSs which were avoided in multiplex panel construction. The ability of the nine multiplexes to detect pathology associated microdeletions is equal to or greater than screening protocols used in other studies. Furthermore, the data suggest a fourth AZF region between AZFb and AZFc, which we have termed AZFd. Patients with microdeletions restricted to AZFd may present with mild oligozoospermia or even normal sperm counts associated with abnormal sperm morphology. Though a definitive genotype/phenotype correlation does not exist, large deletions spanning multiple AZF regions or microdeletions restricted to AZFa usually result in patients with Sertoli Cell Only (SCO) or severe oligozoospermia, whereas microdeletions restricted to AZFb or AZFc can result in patients with phenotypes which range from SCO to moderate oligozoospermia. The panel of nine multiplexed reactions, the Y-deletion Detection System (YDDS), provides a fast, efficient and accurate method of assessing the integrity of the Y-chromosome. To date, this study provides the most extensive screening of a proven fertile male population in tandem with 514 infertile males, derived from three different patient selection protocols.