Clinical value of cell counts and indices in testicular fine needle aspiration cytology in primary infertility: diagnostic performance compared with histology

OBJECTIVE: To compare the diagnostic value of testicular fine needle aspiration (FNA) cytology with that of open biopsy in primary infertility and nonobstructive azospermia or severe oligozoospermia, to evaluate the reliability of percentage cell counts and cell indices. STUDY DESIGN: Thirty patients (21 azospermic and 9 severe oligozoospermic) who had samples for testicular FNA obtained from both testis (mean age = 28.7) and open biopsy were included in the prospective study. Primary infertility, history, complete physical examination, hormonal assay and testicular ultrasound data were evaluated. One case was excluded because of an unsatisfactory result in aspiration cytology. The percentage population of Sertoli cells and spermatogenetic cells, in addition to spermatic index, sertoli cell index and sperm-Sertoli cell indexes, was calculated. The statistical analysis was determined using the paired t test. RESULTS: Progressively increasing values of the Sertoli cell index and progressively decreasing values of the sperm--Sertoli cell index were seen in maturation arrest, hypospermatogenesis and Sertoli cell-only syndrome. The difference between mean counts and indices in normal spermatogenesis and other histologic categories was statistically significant (p < 0.05). CONCLUSION: Percentage cell counts and cell indices in testicular FNA significantly correlate with histological categories. In primary male infertility, testicular FNA can be performed instead of open biopsy.     

Modified testicular expression of stress-associated "readthrough" acetylcholinesterase predicts male infertility.

Male infertility is often attributed to stress. However, the protein or proteins that mediate stress-related infertility are not yet known. Overexpression of the "readthrough" variant of acetylcholinesterase (AChE-R) is involved in the cellular stress response in a variety of mammalian tissues. Here, we report testicular overexpression of AChE-R in heads, but not tails, of postmeiotic spermatozoa from mice subjected to a transient psychological stress compared with age-matched control mice. Transgenic mice overexpressing AChE-R displayed reduced sperm counts, decreased seminal gland weight, and impaired sperm motility compared with age-matched nontransgenic controls. AChE-R was prominent in meiotic phase spermatocytes and in tails, but not heads, of testicular spermatozoa from AChE-R transgenic mice. Head-localized AChE-R was characteristic of human sperm from fertile donors. In contrast, sperm head AChE-R staining was conspicuously reduced in samples from human couples for whom the cause of infertility could not be determined, similar to the pattern found in transgenic mice. These findings indicate AChE-R involvement in impaired sperm quality, which suggests that it is a molecular marker for stress-related infertility.     

The cytologic identification and quantification of testicular cell subtypes. Reproducibility and relation to histologic findings in the diagnosis of male infertility

In testicular imprint smears from 100 infertile men (both testicles), stained using the Pappenheim and Papanicolaou methods, the cell forms (light and dark spermatogonia, primary and secondary spermatocytes, spermatids, spermatozoa and Sertoli cells) were identified and quantified by the counting of 500 consecutive cells. Identification of the cell types, which are described and illustrated, was consistent and reproducible; the advantages of the different stainings in their analysis is documented. The cell counts were tested for reproducibility and compared to the histologic diagnoses and sperm counts. Statistical analysis showed the highest reproducibility for cells frequently encountered in smears (0.99 for Sertoli cells and 0.98 for spermatozoa) and the lowest but still satisfactory reproducibility for rare or arbitrarily defined cell forms (0.71 for dark spermatogonia and 0.76 for secondary spermatocytes). The high reproducibility of the smear quantification permits the introduction of a number of indices, defining clinically useful relations between cell types that are indicative of various types of infertility. The data obtained by cytologic quantification showed reasonably good correlation with the histologic diagnoses of desquamation and focal fibrosis and excellent correlation with Sertoli cells only, arrested spermatogenesis and complete fibrosis. The cytologic quantification of testicular smears adds considerable information to the diagnosis of impaired fertility and should be instituted in properly equipped centers.     

Downregulation of microRNA-383 is associated with male infertility and promotes testicular embryonal carcinoma cell proliferation by targeting IRF1

Our previous studies have shown that microRNA-383 (miR-383) expression is downregulated in the testes of infertile men with maturation arrest (MA). However, the underlying mechanisms of miR-383 involved in the pathogenesis of MA remain unknown. In this study, we showed that downregulation of miR-383 was associated with hyperactive proliferation of germ cells in patients with mixed patterns of MA. Overexpression of miR-383 in NT2 (testicular embryonal carcinoma) cells resulted in suppression of proliferation, G1-phase arrest and induction of apoptosis, whereas silencing of miR-383 reversed these effects. The effects of miR-383 were mediated through targeting a tumor suppressor, interferon regulatory factor-1 (IRF1), and miR-383 was negatively correlated with IRF1 protein expression in vivo. miR-383 inhibited IRF1 by affecting its mRNA stability, which subsequently reduced the levels of the targets of IRF1, namely cyclin D1, CDK2 and p21. Downregulation of IRF1 or cyclin D1, but not that of CDK2, enhanced miR-383-mediated effects, whereas silencing of p21 partially inhibited the effects of miR-383. Moreover, miR-383 downregulated CDK4 by increasing proteasome-dependent degradation of CDK4, which in turn resulted in an inhibition of phosphorylated retinoblastoma protein (pRb) phosphorylation. These results suggest that miR-383 functions as a negative regulator of proliferation by targeting IRF1, in part, through inactivation of the pRb pathway. Abnormal testicular miR-383 expression may potentiate the connections between male infertility and testicular germ cell tumor.     

Role of fine needle aspiration cytology in nonneoplastic testicular and scrotal lesions and male infertility

OBJECTIVE: To study the role of fine needle aspiration (FNA) in male infertility and in nonneoplastic lesions of the testis and scrotum. STUDY DESIGN: In a retrospective study over a 5-year period, 164 cases of FNA of testicular and scrotal nonneoplastic lesions were retrieved. Aspiration was performed with a 23-gauge needle on a 20-mL syringe. RESULTS: Of 164 cases, 27 (16%) remained inconclusive; they were mainly from epididymal lesions. The remaining 137 cases were categorized as inflammatory lesions, 52 (31.7%); noninflammatory lesions, 42 (25.6%); and infertility cases, 43 (26.2%). Among the inflammatory lesions, 33 cases had nonspecific inflammation, 13 had granulomatous epididymoorchitis, 3 cases were of spermatic granuloma, and 3 cases revealed microfilariae. Noninflammatory lesions included 25 cases of spermatocele, 8 of hematoma/torsion, 5 of hydrocele, 3 of benign epididymal cyst and 1 of calcinosis cutis. Among the patients investigated for infertility, 23 (53%) had normal spermatogenesis, 6 (14%) had Sertoli cells only, 5 (119%) had maturation arrest, 6 (14%) showed hypospermatogenesis, and 3 (7%) showed an atrophic pattern. CONCLUSION: FNA of the testis and scrotum is a simple, quick, minimally invasive and painless outpatient procedure. The sample obtained is more representative than biopsy as several separate punctures can be made, and there is no local scarring.     

The testicular form of hormone-sensitive lipase HSLtes confers rescue of male infertility in HSL-deficient mice

Inactivation of the hormone-sensitive lipase gene (HSL) confers male sterility with a major defect in spermatogenesis. Several forms of HSL are expressed in testis. HSLtes mRNA and protein are found in early and elongated spermatids, respectively. The other forms are expressed in diploid germ cells and interstitial cells of the testis. To determine whether the absence of the testis-specific form of HSL, HSLtes, was responsible for the infertility in HSL-null mice, we generated transgenic mice expressing HSLtes under the control of its own promoter. The transgenic animals were crossed with HSL-null mice to produce mice deficient in HSL in nongonadal tissues but expressing HSLtes in haploid germ cells. Cholesteryl ester hydrolase activity was almost completely blunted in HSL-deficient testis. Mice with one allele of the transgene showed an increase in enzymatic activity and a small elevation in the production of spermatozoa. The few fertile hemizygous male mice produced litters of very small to small size. The presence of the two alleles led to a doubling in cholesteryl ester hydrolase activity, which represented 25% of the wild type values associated with a qualitatively normal spermatogenesis and a partial restoration of sperm reserves. The fertility of these mice was totally restored with normal litter sizes. In line with the importance of the esterase activity, HSLtes transgene expression reversed the cholesteryl ester accumulation observed in HSL-null mice. Therefore, expression of HSLtes and cognate cholesteryl ester hydrolase activity leads to a rescue of the infertility observed in HSL-deficient male mice.     

Retroperitoneal and metachronous testicular germ cell tumors with different histology and teratoma growing syndrome--a case report

It is presented a case of a 32-year-old male with the three primary tumors diagnosed within a time period of 3 years; retroperitoneal nonseminoma in 2002, retroperitoneal mature teratoma in 2004, and metachronous testicular seminoma in 2005. We discuss the unusual presentation of these three rare events occurring in the same patient without known risk factors.     

Effect of prenatal treatment with busulfan on the hypothalamo-pituitary axis, genital tract and testicular histology of prepubertal male rats

Female Wistar rats were treated with busulfan or with solvent on Day 20 of pregnancy. Thirty male offspring of each group were killed at 38 days of age. In busulfan-treated rats, compared to controls, hypothalamic LH-RH content was decreased by 52%, whereas pituitary LH and FSH concentrations were increased by 60 and 43% respectively. Plasma LH and FSH were increased by 112 and 275% respectively. Prolactin concentrations were not changed, but plasma testosterone concentration was decreased by 48%. The total number of Leydig cells per testis was decreased by 52%, and LH binding sites per testis were decreased by 70%. The total number of Sertoli cells was decreased by 44%, while FSH binding sites per testis were decreased by 62%. Spermatogenesis was practically absent after prenatal exposure to busulfan. These data demonstrate that on Day 20 of pregnancy all the dividing cells in the fetal testes were depleted by an antimitotic treatment. The stimulation of the hypothalamo-pituitary axis could have been partly induced by the decrease in testosterone production, and by the aplasia of germ cells involving modifications of the remaining Sertoli and Leydig cells.     

Oxidative stress EPR measurement in human liver by radical-probe technique. Correlation with etiology,histology and cell proliferation

The role of reactive oxygen species (ROS) in liver disease is controversial. This mostly reflects the difficulties to quantify ROS in vivo, particularly in humans. We aimed to measure the presence of ROS in diseased human liver and identify possible relations between ROS levels and etiology, histology and hepatocyte proliferation. Liver biopsy specimens from 102 individuals: 18 healthy controls and 84 patients (42 HCV chronic hepatitis (CHC), 19 HBV chronic hepatitis (CHB), 7 PBC, 4 PSC, 4 HCV relapsing hepatitis after liver transplantation, 3 autoimmune hepatitis, 3 hepatocellular carcinoma, 2 alcoholic hepatitis) underwent analysis by radical-probe electron paramagnetic resonance (EPR). ROS in patients (median = 5 x 10(-6) mmol/mg) were higher than in controls (median = 3 x 10(-11) mmol/mg) (p < 0.001). Progressively increasing levels of ROS were recorded passing from control values to CHB (median = 4 x 10(-7) mmol/mg), CHC (median = 3 x 10(-6) mmol/mg) and PBC (median = 2 x 10(-5) mmol/mg), the differences being significant (p < 0.001). ROS in CHC positively correlated with histological disease activity (r = 0.92; p < 0.001). No correlation was found between ROS and hepatocyte proliferation rate, presence/degree of steatosis, serum ferritin levels and aminotransferases. ROS overproduction in liver appears to be a common thread linking different pathologic conditions and seems to be influenced by diseases' etiologies.