Expression of miRNA-371a-3p in seminal plasma and ejaculate is associated with sperm concentration.

BACKGROUND: The microRNAs of the miR-371-3 cluster are novel serum markers for testicular germ cell tumors. Sporadic reports suggested the expression of this miRNA in semen. OBJECTIVES: To verify the expression of miR-371a-3p in seminal plasma and unprocessed ejaculate; to compare seminal plasma miRNA levels in germ cell tumors patients with those of controls; to look for an association of miRNA levels with semen quality. MATERIALS AND METHODS: The miR-371a-3p expression was analyzed with qPCR. The study population consisted of 100 participants: seminal plasma samples from 20 germ cell tumors patients and 30 controls, serum samples from 12 healthy men, ejaculate samples from 38 men undergoing fertility testing. RESULTS: The seminal plasma miR-371a-3p levels of germ cell tumors patients were not different from controls. The miRNA expression was very low in serum but much higher in seminal plasma. In ejaculate samples, the miRNA expression significantly correlated with sperm concentration and the total sperm count. DISCUSSION: miR-371-a-3p is present in sperm-containing fluids. Seminal plasma levels cannot be used to distinguish germ cell tumors from controls. The correlation with sperm concentration in ejaculate samples suggests the spermatozoa as possible source of miR-371a-3p production. CONCLUSION: The miR-371a-3p levels in ejaculate could represent a novel biomarker for the non-invasive evaluation of male infertility.

A molecular rheostat adjusts auxin flux to promote root protophloem differentiation.

Auxin influences plant development through several distinct concentration-dependent effects 1 . In the Arabidopsis root tip, polar auxin transport by PIN-FORMED (PIN) proteins creates a local auxin accumulation that is required for the maintenance of the stem-cell niche2-4. Proximally, stem-cell daughter cells divide repeatedly before they eventually differentiate. This developmental gradient is accompanied by a gradual decrease in auxin levels as cells divide, and subsequently by a gradual increase as the cells differentiate5,6. However, the timing of differentiation is not uniform across cell files. For instance, developing protophloem sieve elements (PPSEs) differentiate as neighbouring cells still divide. Here we show that PPSE differentiation involves local steepening of the post-meristematic auxin gradient. BREVIS RADIX (BRX) and PROTEIN KINASE ASSOCIATED WITH BRX (PAX) are interacting plasma-membrane-associated, polarly localized proteins that co-localize with PIN proteins at the rootward end of developing PPSEs. Both brx and pax mutants display impaired PPSE differentiation. Similar to other AGC-family kinases, PAX activates PIN-mediated auxin efflux, whereas BRX strongly dampens this stimulation. Efficient BRX plasma-membrane localization depends on PAX, but auxin negatively regulates BRX plasma-membrane association and promotes PAX activity. Thus, our data support a model in which BRX and PAX are elements of a molecular rheostat that modulates auxin flux through developing PPSEs, thereby timing PPSE differentiation.

Isolation and gene expression analysis of single potential human spermatogonial stem cells.

STUDY HYPOTHESIS: It is possible to isolate pure populations of single potential human spermatogonial stem cells without somatic contamination for down-stream applications, for example cell culture and gene expression analysis. STUDY FINDING: We isolated pure populations of single potential human spermatogonial stem cells (hSSC) without contaminating somatic cells and analyzed gene expression of these cells via single-cell real-time RT-PCR. WHAT IS KNOWN ALREADY: The isolation of a pure hSSC fraction could enable clinical applications such as fertility preservation for prepubertal boys and in vitro-spermatogenesis. By utilizing largely nonspecific markers for the isolation of spermatogonia (SPG) and hSSC, previously published cell selection methods are not able to deliver pure target cell populations without contamination by testicular somatic cells. However, uniform cell populations free of somatic cells are necessary to guarantee defined growth conditions in cell culture experiments and to prevent unintended stem cell differentiation. Fibroblast growth factor receptor 3 (FGFR3) is a cell surface protein of human undifferentiated A-type SPG and a promising candidate marker for hSSC. It is exclusively expressed in small, non-proliferating subgroups of this spermatogonial cell type together with the pluripotency-associated protein and spermatogonial nuclear marker undifferentiated embryonic cell transcription factor 1 (UTF1). STUDY DESIGN, SAMPLES/MATERIALS, METHODS: We specifically selected the FGFR3-positive spermatogonial subpopulation from two 30 mg biopsies per patient from a total of 37 patients with full spermatogenesis and three patients with meiotic arrest. We then employed cell selection with magnetic beads in combination with a fluorescence-activated cell sorter antibody directed against human FGFR3 to tag and visually identify human FGFR3-positive spermatogonia. Positively selected and bead-labeled cells were subsequently picked with a micromanipulator. Analysis of the isolated cells was carried out by single-cell real-time RT-PCR, real-time RT-PCR, immunocytochemistry and live/dead staining. MAIN RESULTS AND THE ROLE OF CHANCE: Single-cell real-time RT-PCR and real-time RT-PCR of pooled cells indicate that bead-labeled single cells express FGFR3 with high heterogeneity at the mRNA level, while bead-unlabeled cells lack FGFR3 mRNA. Furthermore, isolated cells exhibit strong immunocytochemical staining for the stem cell factor UTF1 and are viable. LIMITATIONS, REASONS FOR CAUTION: The cell population isolated in this study has to be tested for their potential stem cell characteristics via xenotransplantation. Due to the small amount of the isolated cells, propagation by cell culture will be essential. Other potential hSSC without FGFR3 surface expression will not be captured with the provided experimental design. WIDER IMPLICATIONS OF THE FINDINGS: The technical approach as developed in this work could encourage the scientific community to test other established or novel hSSC markers on single SPG that present with potential stem cell-like features. STUDY FUNDING AND COMPETING INTERESTS: The project was funded by the DFG Research Unit FOR1041 Germ cell potential (SCH 587/3-2) and DFG grants to K.v.K. (KO 4769/2-1) and A.-N.S. (SP 721/4-1). The authors declare no competing interests.

[Evaluation of obstructive interval, presence of sperm granulomas and patient age as predictors of spermatogenesis in a cohort of men undergoing vasectomy reversal]. / Evaluation von Vasektomieintervall, Spermagranulom und Patientenalter als Prädiktoren der spermatogenetischen Aktivität vasektomierter Männer.

INTRODUCTION: For more than 50 years vasectomy reversal is a routinely performed procedure in the field of urology. However, there is no scientific agreement about morphological changes of the testes caused by vasectomy. The existing evidence for reduced fertility rates following vasectomy reversal demands a clear statement regarding potential histological changes and impaired spermatogenesis following vasectomy. Thus far there is little knowledge about potential histological changes of the testis caused by vasectomy. MATERIAL AND METHODS: 330 consecutive patients who underwent vasectomy reversal had bilateral testicular biopsies which were evaluated utilising semi-thin sections. The number of mature spermatids per seminiferous tubule was considered the variable of interest as it represents an objective and reproducible parameter of spermatogenesis. The number of mature spermatids per tubule was correlated with patient age, obstructive interval and the presence or absence of sperm granulomas via the chi-square test. RESULTS: Overall, 570 sections of 285 patients were eligible for evaluation. The mean patient age was 41.2 years (range 27-63 years, SD +/- 6.5 years) with a mean obstructive interval of 105.9 months (range 12-328, SD +/- 66.1). 56 patients (19.6 %) had a sperm granuloma on the right and 22 (11.6 %) on the left ductus deferens. There was no statistically significant correlation between the presence of a sperm granuloma with the number of mature spermatids per tubule (p = 0.717). Furthermore, there was neither an association of obstructive interval (p = 0.144) nor patient age (p = 0.168) with spermatogenesis. CONCLUSION: Regular spermatogenetic activity in all examined samples with development of mature spermatids was shown. Furthermore, for the first time we were able to demonstrate in a large cohort of patients that neither patient age nor obstructive interval nor sperm granuloma have a significant impact on spermatogenesis.

A new paradigm for profiling testicular gene expression during normal and disturbed human spermatogenesis.

The aim of this study was to identify gene expression patterns of the testis that correlate with the appearance of distinct stages of male germ cells. We avoided the pitfalls of mixed pathological phenotypes of the testis and circumvented the inapplicability of using the first spermatogenic wave as done previously on rodents. This was accomplished by using 28 samples showing defined and highly homogeneous pathologies selected from 578 testicular biopsies obtained from 289 men with azoospermia (two biopsies each). The molecular signature of the different developmental stages correlated with the morphological preclassification of the testicular biopsies, as shown by resampling-based hierarchical clustering using different measures of variability. By using analysis of variance (ANOVA) and extensive permutation analysis, we filtered 1181 genes that exhibit exceptional statistical significance in testicular expression and grouped subsets with transcriptional changes within the pre-meiotic (348 genes), post-meiotic (81 genes) and terminal differentiation (38 genes) phase. Several distinct molecular classes, metabolic pathways and transcription factor binding sites are involved, depending on the transcriptional profile of the gene clusters that were built using a novel clustering procedure based on not only similarity but also statistical significance.

Multicolor fluorescence in situ hybridization (M-FISH) on cells from urine for the detection of bladder cancer.

Bladder cancer is the fifth most common cancer in adults. Because of the high recurrence rate (up to 70%) new tumor markers for urine are necessary for monitoring patients. In this study, we investigated the value of M-FISH on cells from urine for the detection of bladder cancer. Urine samples from 141 patients suspicious of bladder cancer were analyzed in this study. Cells were isolated from urine before surgical therapy. For FISH analysis, a commercial kit (UroVysion) containing hybridization probes for chromosomes 3, 7, 9p21 and 17, was used. Twenty-five cells were analyzed in each case by two observers. A FISH result was obtained in 121 cases. Overall, sensitivity was 60% and specificity reached 82.6%. Sensitivity and specificity by cytology were 24.1% and 90.5%, respectively. Analyzing results concerning T-category, sensitivity of FISH and cytology was 36.1% and 15% in pTa, 65.2 and 25.7% in pT1, 100% and 66.7% in pT2-3 tumors, respectively. Concerning tumor grade, similar results were obtained: sensitivity was 37% and 14% in G1, 65.4% and 40% in G2, 91.7% and 50% in G3 tumors, for FISH and cytology, respectively. In conclusion, FISH on cells from urine has been shown in all studies to be highly sensitive and specific for detection of bladder cancer. Sensitivity of FISH is higher than conventional cytology and can be used in routine diagnosis additionally to conventional cytology especially in doubtful or negative cases. FISH can detect recurrence earlier than other methods like cytology, cystoscopy or biopsy histological examination.

Megalospermatocytes in the human testis exhibit asynapsis of chromosomes.

Testis biopsies of three infertile patients were identified, which showed a predomination of megalospermatocytes in the seminiferous tubules. Megalospermatocytes are very large primary spermatocytes indicating a spermatogenic arrest. Because of the high percentage of these germ cells it was possible to apply a whole-mount spreading technique to investigate the chromosomal pairing behaviour in prophase I of meiosis. It could be shown that most of the megalospermatocytes exhibited extensive chromosomal asynapsis, suggesting that a characteristic meiotic disorder may give rise to reduced fertility, or even infertility.

[The role of the tibiofibular syndesmotic and the deltoid ligaments in stabilizing Weber B type ankle joint fractures--an experimental investigation]. / Beitrag der Syndesmosen und des Deltabandes zur Stabilität der Aussenknöchelfraktur Typ Weber B. Eine experimentelle Untersuchung.

The purpose of the present biomechanical investigation was to check the functional importance of the syndesmosis ligaments and of the deltoid ligament for ankle fracture type B according to the AO-Weber classification. We constructed a special fixation clamp, with 12 fresh and unembalmed lower legs being tested for lateral shift (mm) and ten for tibiotalar rotation. All specimens were exposed in the same neutral position. Transverse loads (F(y)) varied between 0 and 150 N, axial loads (F(z)) between 0, 300, 600 and 1,000 N and rotational loads (F(r)) between 2.4 and 4.9 Nm. All series were repeated according to supination-eversion (SE) injury patterns of the Lauge-Hansen classification. Syndesmotic ligaments and the fibula were incrementally sectioned from anterior to posterior. Type SE I consisted of an isolated incision of the anterior syndesmosis ligament. Type SE II had an additional oblique fracture of the fibula at the height of the tibiofibular syndesmosis. In type SE III injuries, in addition to the fibular fracture, a complete rupture of the syndesmosis ligaments was present, and for type SE IV lesions the deltoid ligaments were incised. The transverse load-displacement curve was s-shaped in all uninjured joints,with the highest gradient between 10 and 20 N with no axial compression. Without axial compression in cases of F(y)=25 N transverse loads, the mean talus translation was 0.51 mm. Following type II injuries, the average talus translation was 0.68 mm (not significant) and rose to an average of 0.95 mm ( P <0.01) in type III injuries. After additional incision of the deltoid ligaments, the ankle joint subluxed permanently when more than 5-10 N transverse loads were applied. Axial loads of 300 N or more resulted in a considerable reduction in talus translations, indicating increased stability and congruency within the joint complex. In this way, the vertical loading of the ankle joints always contributed to joint stability. The average internal tibiotalar rotation reached with a torque of 2.4 Nm was 3.52 degrees and with 4.9 Nm 5.15 degrees when no axial compression was applied. External rotation measured -6.36 degrees and -8.62 degrees, respectively. Following the experimental protocol, significant increases were noted for external rotation at SE II degrees injuries ( P =0.003) and for internal rotation at SE III degrees ( P =0.03) injuries. Our data support the proposition that the deltoid ligaments and the posterior syndesmosis play a key role in the stability of ankle fractures for supination-eversion injuries. If these structures remain intact, conservative and early functional treatment are recommended in patients with minimal (<2 mm) or no fracture displacement. This concept is confirmed by the literature dealing with clinical mid- and long-term follow-up studies.

Injury prophylaxis in paragliding.

OBJECTIVES: To show trends in paragliding injuries and derive recommendations for safety precautions for paraglider pilots on the basis of accident statistics, interviews, questionnaires, medical reports, and current stage of development of paragliding equipment. METHODS: All paragliding accidents in Germany have to be reported. Information on 409 accidents was collected and analysed for the period 1997-1999. RESULTS: There was a substantial decrease in reported accidents (166 in 1997; 127 in 1998; 116 in 1999). The number of accidents resulting in spinal injuries was 62 in 1997, 42 in 1998, and 38 in 1999. The most common cause of accident was deflation of the glider (32.5%), followed by oversteering (13.9%), collision with obstacles (12.0%), take off errors (10.3%), landing errors (13.7%), misjudgment of weather conditions (4.9%), unsatisfactory preflight checks (4.9%), mid-air collisions with other flyers (2.2%), accidents during winching (2.2%), and defective equipment (0.5%). Accidents predominantly occurred in mountain areas. Fewer than 100 flights had been logged for 40% of injured pilots. In a total of 39 accidents in which emergency parachutes were used, 10 pilots were seriously injured (26%) and an additional three were killed (8%). CONCLUSIONS: Injuries in paragliding caused by unpredictable situations can be minimised by (a) using safer gliders in the beginner or intermediate category, (b) improving protection systems, such as padded back protection, and (c) improving pilot skills through performance and safety training.

[Prognostic factors for avascular necrosis following talar fractures]. / Prognostische Faktoren für die avaskuläre Nekrose nach Talusfrakturen.

AIM: We performed an investigation of factors for avascular necroses after talus fracture and on the reliability of the Hawkins Sign. METHOD: From 1984 until 1997 a total of 98 patients with 99 talus fractures were surgically treated. Of these, 79 patients with 80 fractures were examined clinically and radiologically. The average postoperative interval was 6 years and 2 months. RESULTS: With respect to the 65 central fractures, the rate of necrosis amounted to 14 %, that of collum fractures to 17 %. Necroses arose solely in dislocated central fractures of the talus, type III and IV according to Marti/Weber fracture classification. The rate of necrosis rose with the degree of dislocation of the fractures. In 24 patients the Hawkins Sign could be retrospectively investigated. It proved to be a relatively reliable sign for vitality since only 1 out of 12 patients with positive or partial positive Hawkins Sign developed avascular necrosis. Neither a short interval between accident and operation, the age at the time of the accident, nor the ipsilateral fracture of the medial malleolus showed a necrosis preventive influence. In 5 out of 9 talus necroses the patients were very or mostly satisfied with the result of their treatment. CONCLUSION: The Hawkins Sign proved to be a relatively reliable sign for vitality of the talus after fracture. Risk for avascular necrosis increases according to the degree of fracture dislocation.

Quantification of telomerase activity, porphobilinogen deaminase and human telomerase reverse transcriptase mRNA in testicular tissue - new parameters for a molecular diagnostic classification of spermatogenesis disorders.

The aim of the study was to evaluate the quantitative detection of human telomerase reverse transcriptase (hTERT) mRNA and telomerase activity as new molecular diagnostic parameters for a subclassification of spermatogenesis disorders. Telomerase activity was detected by a quantitative telomerase PCR ELISA, and hTERT mRNA expression was quantified by fluorescence real-time RT-PCR in a LightCycler in cryopreserved testicular tissue specimens. This was paralleled by a histological workup. The discriminant analysis showed that detection of normalized hTERT expression was able to correctly classify 89.0% of the investigated tissue specimens into the subgroups of full spermatogenesis, maturation arrest or Sertoli-cell-only syndrome. In contrast, discriminant analysis revealed an only 58% accuracy of telomerase activity for the investigated tissue specimens. This study shows that the quantification of hTERT expression in testicular tissue by real-time fluorescence RT-PCR is well suited for correctly classifying spermatogenesis disorders and proved to be markedly superior to the determination of telomerase activity.

Quantification of the expression level of the gene encoding the catalytic subunit of telomerase in testicular tissue specimens predicts successful sperm recovery.

BACKGROUND: The objective of the present study was to evaluate the quantitative detection of human telomerase reverse transcriptase (hTERT) mRNA as a new molecular diagnostic parameter in the work-up of testicular tissue specimens from patients presenting with non-obstructive azoospermia. M ETHODS: hTERT mRNA expression was quantified in 49 cryopreserved testicular tissue specimens by fluorescence real-time RT-PCR in a LightCycler. This was paralleled by conventional histological work-up in all tissue specimens and additional semithin sectioning preparation in cases with maturation arrest (n = 20) and Sertoli cell-only syndrome (SCOS; n = 12). RESULTS: The average normalized hTERT expression (N(hTERT)) was 136.1 +/- 41.7 copies (mean +/- standard deviation) in tissue specimens with presence of haploid germs cells, N(hTERT) = 48.2 +/- 21.0 copies in those with maturation arrest and N(hTERT) = 2.7 +/- 2.8 copies in those with SCOS. The discriminant analysis showed that detection of N(hTERT) was able correctly to classify 89.0% of the investigated tissue specimens. CONCLUSIONS: Our results demonstrate that quantitative detection of hTERT mRNA expression in testicular tissue enables a molecular-diagnostic subclassification of spermatogenesis disorders. Quantitative detection of hTERT in testicular biopsies is thus well suited for predicting successful sperm recovery in patients with azoospermia and is a useful molecular diagnostic parameter for supplementing the histopathological evaluation.

Effects of various cryopreservation media and freezing-thawing on the morphology of rat testicular biopsies.

Currently, testicular sperm extraction is successfully combined with intracytoplasmic sperm injection into the oocyte (ICSI). Several pieces of a testicular biopsy can be frozen and thawed until the ICSI attempt. In this study, the effects of freezing-thawing on the morphology of rat testicular biopsies stored in different cryopreservation media were analysed. Each cryopreservation medium contained glycerol and/or dimethyl sulfoxide (DMSO) as cryoprotectants. In general, both glycerol and DMSO, when applied at moderate concentrations (6-25%), preserved the structure of the seminiferous epithelium. The freezing-thawing procedure had no significant effect on tubular diameter; however, it caused a 'folding' of the lamina propria and notable damage to Sertoli cells, spermatogonia and spermatocytes. Round and elongated spermatids and spermatozoa displayed occasional nuclear damage, vacuolization, and shrinkage/swelling of the cytoplasm. However, the vast majority of these cells maintained their normal structure in nearly all the applied cryomedia. It is concluded that freezing-thawing of testicular biopsies, and the cryopreservation medium, have a significant impact on the structure of the seminiferous epithelium, particularly on its basal compartment.

Phonotaxis during walking and flight: are differences in selectivity due to predation pressure?

Female selectivity was tested in Tettigonia viridissima during two different phonotaxis situations; compensated walking and tethered flight. For two of the three temporal parameters that are important for call recognition in T. viridissima, selectivity was similar in the two situations. Selectivity for the third parameter (minimum interval duration between the double pulses) was much higher during walking than during flight: walking females responded only to stimuli with intervals of 28 ms or longer, while call models with intervals of 18 ms were attractive during flight. One interneuron (TN-1) is probably involved in filtering the minimum interval duration. As this neuron is also the most likely candidate for transmitting bat calls during flight, it is suggested that the selectivity differences between walking and flying might be due to the need for detecting predator signals during flight, when TN-1 would be occupied listening for bats. With TN-1 unavailable for song processing during flight, temporal selectivity for the minimum interval duration should be reduced, as was found here.

Expression of alpha1-adrenergic receptor subtypes in heart cell culture.

Three alpha1-AR subtypes have been cloned so far and are designated as alpha1a, alpha1b,, and alpha1d. Organ-specific distribution pattern and subtype-specific effects are known but not fully understood. To address a cell-type specific expression pattern in the heart we investigated expression pattern of alpha1-AR subtypes on RNA- and protein-level in heart tissue, cultured cardiomyocytes and non-myocytes of the rat. Each alpha1AR-subtype mRNA was present in neonatal and adult rat heart culture but the relative distribution pattern was significantly different. While the alpha1a-AR subtype is preferentially expressed in adult cardiomyocytes, the alpha1b-AR subtype was preferentially expressed in the non-myocyte cell fraction. The RT-PCR results were confirmed by Western-blotting (alpha1b) and immunocytochemical studies. Incubation with an alpha1-agonist (phenylephrine) for 72 h led to a significant reduction of the alpha1b-AR in neonatal heart cell culture on both mRNA and protein level. In contrast, incubation with an alpha1-antagonist (prazosin) induced a 1.6 fold upregulation of the alpha1a-AR mRNA without significant effects on radioligand binding and functional assay. The results indicate a distribution pattern of the alpha1-AR subtype which is specific for cell type and ontogeny of the rat heart and may be regulated by adrenergic agents.

[Clinical pedographic radiological findings after basal valgus osteotomy of metatarsal 1--a prospective study]. / Klinisch-pedographischradiologische Befunde nach valgisierender Basisosteotomie des Metatarsale 1--eine prospektive Studie.

AIM: This study investigated the influence of valgusizing base wedge osteotomy of metatarsal 1 (MT 1) subsequently on the entire forefoot. METHOD: Pre- and post-operatively 22 cases were investigated between 1998 and 2000 both clinically and radiologically with pedographic analysis in 20 of these cases. RESULTS: According to the Kitaoka Score we could establish preoperative 37 and postoperative 72 out of 100 possible points. Following the MT 1-base wedge osteotomy with a distal soft-tissue procedure, the central forefoot region (MT 2/3) showed reduced pressure-induced pain, unchanged callus development, and reduced maximum load pressure. When the Plus-Index (MT 1 longer than MT 2) could be preserved in the valugusizing MT 1-base wege osteotomy, the metatarsalgia in the central forefoot region not only improved generally, but also the maximum loading pressure was clearly reduced. In cases with a post-operative Minus-Index (MT 1 shorter than MT 2), as well as in those Plus-Index cases undergoing a Keller procedure in the valgusizing base wedge osteotomy, the elevated pressure values in the central forefoot region persisted. CONCLUSION: In valgusizing MT 1-base wege osteotomy with preservation of the Plus-Index (MT 1 longer MT 2), metatarsalgia can be improved and the maximum loading pressure in the central forefoot can be reduced.

High deletion frequency of the complete AZFa sequence in men with Sertoli-cell-only syndrome.

We have developed a rapid screening protocol for deletion analysis of the complete AZFa sequence (i.e. 792 kb) on the Y chromosome of patients with idiopathic Sertoli-cell-only (SCO) syndrome. This Y deletion was mapped earlier in proximal Yq11 and first found in the Y chromosome of the SCO patient JOLAR, now designated as the AZFa reference patient. We now show that similar AZFa deletions occur with a frequency of 9% in the SCO patient group. In two multiplex polymerase chain reaction experiments, deletions of the complete AZFa sequence were identified by a typical deletion pattern of four new sequence-tagged sites (STS): AZFa-prox1, positive; AZFa-prox2, negative; AZFa-dist1, negative; AZFa-dist2, positive. The STS were established in the proximal and distal neighbourhoods of the two retroviral sequence blocks (HERV15yq1 and HERV15yq2) which encompass the break-point sites for AZFa deletions of the human Y chromosome. We have found deletions of the complete AZFa sequence always associated with a uniform SCO pattern on testicular biopsies. Patients with other testicular histologies as described in the literature and in this paper have only partial AZFa deletions. The current AZFa screening protocols can therefore be improved by analysing the extension of AZFa deletions. This may provide a valuable prognostic tool for infertility clinics performing testicular sperm extraction, as it would enable the exclusion of AZFa patients with a complete SCO syndrome.