Matthias Trottmann

A Nonsense Mutation in TMEM95 Encoding a Nondescript Transmembrane Protein Causes Idiopathic Male Subfertility in Cattle

Genetic variants underlying reduced male reproductive performance have been identified in humans and model organisms, most of them compromising semen quality. Occasionally, male fertility is severely compromised although semen analysis remains without any apparent pathological findings (i.e., idiopathic subfertility). Artificial insemination (AI) in most cattle populations requires close examination of all ejaculates before insemination. Although anomalous ejaculates are rejected, insemination success varies considerably among AI bulls. In an attempt to identify genetic causes of such variation, we undertook a genome-wide association study (GWAS). Imputed genotypes of 652,856 SNPs were available for 7962 AI bulls of the Fleckvieh (FV) population. Male reproductive ability (MRA) was assessed based on 15.3 million artificial inseminations. The GWAS uncovered a strong association signal on bovine chromosome 19 (P = 4.08×10−59). Subsequent autozygosity mapping revealed a common 1386 kb segment of extended homozygosity in 40 bulls with exceptionally poor reproductive performance. Only 1.7% of 35,671 inseminations with semen samples of those bulls were successful. None of the bulls with normal reproductive performance was homozygous, indicating recessive inheritance. Exploiting whole-genome re-sequencing data of 43 animals revealed a candidate causal nonsense mutation (rs378652941, c.483C>A, p.Cys161X) in the transmembrane protein 95 encoding gene TMEM95 which was subsequently validated in 1990 AI bulls. Immunohistochemical investigations evidenced that TMEM95 is located at the surface of spermatozoa of fertile animals whereas it is absent in spermatozoa of subfertile animals. These findings imply that integrity of TMEM95 is required for an undisturbed fertilisation. Our results demonstrate that deficiency of TMEM95 severely compromises male reproductive performance in cattle and reveal for the first time a phenotypic effect associated with genomic variation in TMEM95.

Risk factors for pelvic lymphoceles post-radical prostatectomy.

Background:  Lymphoceles (LC) represent a well‐described rare complication post‐radical prostatectomy (RP). Our aim was to determine risk factors and to develop possible prevention strategies for LC in a community‐based study.

Acoustic radiation force impulse imaging of the testes: A preliminary experience

PURPOSE Acoustic radiation force impulse imaging (ARFI) is a newly developed technique for the sonographic quantification of tissue elasticity. It has been used in the assessment of various abdominal organs. The testes are extraabdominal organs with limited overlying tissue, making them ideally suited to ARFI assessment. To our knowledge no published data exist on ARFI elastography of the testes. METHOD 23 male volunteers (mean age 45.13 ± 17.3, range 23-75) with no known testicular pathology underwent normal B-mode sonography with ARFI elastometry of both testes using a Siemens Acuson S2000™ (Siemens Healthcare, Germany) system. 15 measurements were performed on each testis; 5 each in the upper pole, middle portion and lower pole. Results were statistically evaluated. RESULTS 95% of the SWV values were found to lie within the reference interval ranging from 0.62 to 1.01 m/s. There was a significant association between measured SWV and age (P = 0.0056) and testicular volume (P = 0.0003). CONCLUSION ARFI elastometry proved to be feasible in the assessment of testicular stiffness. Older age and lower testicular volumes were associated with increased parenchymal stiffness. Further studies with more subjects may be required to define the normal range of values for each age group.

The role of VTIQ as a new tissue strain analytics measurement technique in testicular lesions.

PURPOSE Virtual touch tissue imaging quantification (VTIQ) is a newly developed technique for the sonographic quantification of tissue elasticity. It has been used in the assessment of breast lesions. The purpose of this study was to determine the diagnostic performance of VTIQ in indeterminate testicular lesions. METHODS Twenty patients with known testicular pathology underwent conventional B-mode sonography with additional VTIQ of the testicular lesions using a Siemens Acuson S2000™ and S3000™ (Siemens Medical Solutions, Mountain View, CA, USA) system. Tissue mechanical properties were analysed in the VTIQ examination. The pathologic diagnosis was established after surgery or in the follow-up examination for suspected benign lesions. RESULTS Over 36 months, 22 focal testicular lesions (median lesion size, 18 mm; range, 4-36 mm in 20 patients (median age, 43 years; range, 22-81 years) were examined. Lesions were hyperechoic (n = 1), hypoechoic (n = 14), isoechoic (n = 1), of mixed echogenicity (n = 3) or anechoic (n = 3). Histological examination showed one benign lesion (6.25%) with a mean size of 7 mm and 15 malignant lesions (93.75%) with a mean size of 20 mm. Mean shear wave velocity for normal testicular tissue was 1.17 m/s. No shear wave velocity could be measured in cystic lesions. The rest of the benign lesions showed a mean shear wave velocity of 2.37 m/s. The value of the shear wave velocity in germ cell tumours showed a mean shear wave velocity of 1.94 m/s and for seminoma it showed a mean shear wave velocity of 2.42 m/s. CONCLUSIONS VTIQ is a reliable new method for measuring qualitative and quantitative stiffness of testicular lesions and tissue. The qualitative shear-wave elastography features were highly reproducible and showed good diagnostic performance in unclear testicular lesions. The VTIQ technique is also useful in assessing small testicular nodules and pseudolesions.

Injuries of the renal pelvis and ureter. Diagnosis and management

ZusammenfassungEtwa 5% aller Verletzungen des Harntrakts betreffen das Nierenbecken und den Ureter und stellen eine schwerwiegende Komplikation dar. Mit 75% ist der größere Teil dieser Verletzungen iatrogen bedingt, während nur etwa 25% auf Grund eines stumpfen Bauchtraumas oder einer offenen Verletzung entstehen. Zur Vermeidung von Komplikationen und Verbesserung der Prognose ist eine sofortige Diagnostik und Therapie entscheidend. Die Aussagefähigkeit der präoperativen Diagnosemöglichkeiten ist oft gering, sodass die Verletzung häufig erst bei einer explorativen Laparotomie entdeckt wird. Das Management von Läsionen des oberen Harntrakts ist abhängig von der Schwere und Lokalisation der Verletzung, wobei das oberste Ziel immer der Nierenerhalt sein muss. Grundsätzlich ist die Ureterstentung bei kleinen Verletzungen meist ausreichend und nur bei größeren Traumata eine offene Rekonstruktion indiziert. Hierbei konnte in Langzeitstudien eine hohe Rekonvaleszenz bei zeitnaher und adäquater Therapie dokumentiert werden.AbstractAbout 5% of injuries of the urinary tract affect the renal pelvis and ureter and constitute a severe complication. Around 75% of these injuries are iatrogenic and only about 25% are caused by blunt abdominal trauma or perforation. To avoid complications and improve prognosis, immediate diagnosis and therapy are essential. The diagnostic accuracy of preoperative studies is low, therefore frequently injuries are detected during explorative laparotomy. The management of upper urinary tract lesions depends on severity and localization, whereas the ultimate ambition should always be the preservation of the kidney. As a basic rule, ureteral stenting is mostly sufficient for small lesions, and only larger injuries require open reconstructive techniques. Longitudinal studies document a high degree of functional reconstitution if adequate and immediate treatment is carried out.

Effects of Elevated β-Estradiol Levels on the Functional Morphology of the Testis - New Insights

Elevated estradiol levels are correlated with male infertility. Causes of hyperestrogenism include diseases of the adrenal cortex, testis or medications affecting the hypothalamus-pituitary-gonadal axis. The aim of our study was to elucidate the effects of estradiol treatment on testicular cellular morphology and function, with reference to the treatment regimen received. Testes samples (n = 9) were obtained post-orchiectomy from male-to-female transsexuals within the age range of 26–52 years. Each patient had a minimum of 1–6 years estradiol treatment. For comparison, additional samples were obtained from microscopically unaltered testicular tissue surrounding tumors (n = 7). The tissues obtained were investigated by stereomicroscopy, histochemistry, scanning electron microscopy (SEM) and immunohistochemistry. Our studies revealed that estradiol treatment significantly decreased the diameter of the seminiferous tubules (p < 0.05) and induced fatty degeneration in the surrounding connective tissue. An increase in collagen fiber synthesis in the extracellular matrix (ECM) surrounding the seminiferous tubules was also induced. Spermatogenesis was impaired resulting in mainly spermatogonia being present. Sertoli cells revealed diminished expression of estrogen receptor alpha (ERα). Both Sertoli and Leydig cells showed morphological alterations and glycoprotein accumulations. These results demonstrate that increased estradiol levels drastically impact the human testis.

Shear wave elastography of the testes in a healthy study collective - Differences in standard values between ARFI and VTIQ techniques

INTRODUCTION Shear wave elastography (SWE) is a recent technique in the assessment of tissue elasticity. Different elastography techniques have been described over the years.Acoustic Radiation Force Impulse Imaging (ARFI) uses mechanical excitation of tissue to create detectable shear waves, a higher shear wave velocity being associated with an increased tissue stiffness.The Virtual Touch Tissue Imaging Quantification (VTIQ) method uses a mechanical push pulse as well, additionally creating a colour-coded map, in which tissue stiffness can be measured within a stored map on the ultrasound device after measurement.ARFI has been used in determining standard values in testes of a healthy study collective, VTIQ has already been used in the evaluation of unclear scrotal masses.Both techniques allow an operator-independent examination without application of mechanical pressure. The aim of this study was to determine whether there is a statistically significant difference between shear wave velocity values of both techniques in a healthy collective. MATERIAL AND METHODS Twenty patients without known testicular pathology underwent standard B-mode sonography and additional shear wave elastography in the ARFI mode as well as the VTIQ technique of both testes using the Siemens Acuson S2000™ and S3000™ ultrasound devices (Siemens HealthCare, Erlangen, Germany).Measurements of shear wave velocity were performed in the upper pole, the central portion and the lower pole separately for each testis. Values were described in m/s.Statistical evaluation was performed using paired t-test analysis. RESULTS We measured a mean shear wave velocity of 0.81 m/s using ARFI and 1.07 m/s with VTIQ.Shear wave velocities determined by VTIQ were all significantly higher than values gained in the ARFI mode. (p < 0.001 to p = 0.007). Values were between 0.22 and 0.29 m/s higher, when the examination was performed using VTIQ. CONCLUSION ARFI and VTIQ elastography modes both proved to be feasible techniques in the assessment of testicular tissue elasticity. Consideration of higher values for VTIQ is important, when different elastography measurement results are compared, especially for the application of devices in a clinical setting, e.g. in the work-up of scrotal masses.A calculable factor for a comparison between both devices is desirable, but to be further assessed in largerstudies.

Management of pelvic lymphoceles after radical prostatectomy: A multicentre community based study

IntroductionPelvic lymphoceles (LC) following radical prostatectomy (LC-RP) have an incidence up to 27%. LC-managements constitute 50% of surgical interventions performed in post-RP patients.ObjectivesTo describe a therapeutic algorithm for LC-managements based on a community based representative retrospective study.Patients and methods: Multicentre data from 304 patients with LC-RP were retrospectively examined for LC-managements. RPs were performed by various surgeons from 67 urological departments. All patients had undergone 3 weeks rehabilitation in a specialized hospital where the data base was generated. Indications and results of therapeutic manoeuvres were used to develop a general concept for planning therapy decisions.ResultsMedian age was 64 years. Complications occurred in 9% (28/304) of patients. Median LC-volume was 36 ml (range 20-1800 ml). There were more complications for LCs with ≥100 ml volume than those < 100 ml (27% versus 17%, p = 0.346). Conservative therapy was the standard in uncomplicated cases (87%, 239 of 276 patients), while intervention was done in 13% (puncture and/or drainage, surgery). Surgical intervention was performed significantly more often in complicated cases (82%, 23 from 28 patients; p < 0.001). Based on these data, LCs can be stratified into 3 groups depending on the size and clinical presentation. Therapeutic decisions were used to develop the illustrated new therapy algorithm.ConclusionsThis study based treatment algorithm provides a rationale approach with an accurate LC-classification as regard the indications and decision making for the available LC-RP-therapies. This could facilitate management decisions. Evaluation of this concept prospectively in large patient cohort is mandatory.

Symphysitis following transrectal biopsy of the prostate.

Abstract  Transrectal ultrasound‐guided needle biopsy of the prostate is a widely accepted technique to obtain prostatic tissue for histological examination. Severe complications are rarely seen. We report a case of symphysitis causing hospitalization and severe pain and discomfort of the patient. Possible etiologic factors are traumatic osseous lesions and transport of rectal bacteria to the periosseous region. Especially in small prostates, care should be taken to avoid this condition. Prolonged perioperative antibiotic prophylaxis is mandatory.

Probe-based confocal laser endomicroscopy (pCLE) – a new imaging technique for in situ localization of spermatozoa

In azoospermic patients, spermatozoa are routinely obtained by testicular sperm extraction (TESE). However, success rates of this technique are moderate, because the site of excision of testicular tissue is determined arbitrarily. Therefore the aim of this study was to establish probe-based laser endomicroscopy (pCLE) a noval biomedical imaging technique, which provides the opportunity of non-invasive, real-time visualisation of tissue at histological resolution. Using pCLE we clearly visualized longitudinal and horizontal views of the tubuli seminiferi contorti and localized vital spermatozoa. Obtained images and real-time videos were subsequently compared with confocal laser scanning microscopy (CLSM) of spermatozoa and tissues, respectively. Comparative visualization of single native Confocal laser scanning microscopy (CLSM, left) and probe-based laser endomicroscopy (pCLE, right) using Pro Flex(TM) UltraMini O after staining with acriflavine.