The initial maturation status of marmoset testicular tissues has an impact on germ cell maintenance and somatic cell response in tissue fragment culture.

Successful in vitro spermatogenesis was reported using immature mouse testicular tissues in a fragment culture approach, raising hopes that this method could also be applied for fertility preservation in humans. Although maintaining immature human testicular tissue fragments in culture is feasible for an extended period, it remains unknown whether germ cell survival and the somatic cell response depend on the differentiation status of tissue. Employing the marmoset monkey (Callithrix jacchus), we aimed to assess whether the maturation status of prepubertal and peri-/pubertal testicular tissues influence the outcome of testis fragment culture. Testicular tissue fragments from 4- and 8-month-old (n = 3, each) marmosets were cultured and evaluated after 0, 7, 14, 28 and 42 days. Immunohistochemistry was performed for identification and quantification of germ cells (melanoma-associated antigen 4) and Sertoli cell maturation status (anti-Müllerian hormone: AMH). During testis fragment culture, spermatogonial numbers were significantly reduced (P < 0.05) in the 4- but not 8-month-old monkeys, at Day 0 versus Day 42 of culture. Moreover, while Sertoli cells from 4-month-old monkeys maintained an immature phenotype (i.e. AMH expression) during culture, AMH expression was regained in two of the 8-month-old monkeys. Interestingly, progression of differentiation to later meiotic stage was solely observed in one 8-month-old marmoset, which was at an intermediate state regarding germ cell content, with gonocytes as well as spermatocytes present, as well as Sertoli cell maturation status. Although species-specific differences might influence the outcome of testis fragment experiments in vitro, our study demonstrated that the developmental status of the testicular tissues needs to be considered as it seems to be decisive for germ cell maintenance, somatic cell response and possibly the differentiation potential.

[The antagonistic function of the heart muscle sustains the autoregulation according to Frank and Starling : Part I: Structure and function of heart muscle]. / Die antagonistische Funktion des Herzmuskels unterstützt die Autoregulation nach Frank-Starling : Teil I: Struktur und Funktion des Herzmuskels.

In the tradition of Harvey and according to Otto Frank the heart muscle structure is arranged in a strictly tangential fashion hence all contractile forces act in the direction of ventricular ejection. In contrast, morphology confirms that the heart consists of a 3-dimensional network of muscle fibers with up to two fifths of the chains of aggregated myocytes deviating from a tangential alignment at variable angles. Accordingly, the myocardial systolic forces contain, in addition to a constrictive also a (albeit smaller) radially acting component. Using needle force probes we have correspondingly measured an unloading type of force in a tangential direction and an auxotonic type in dilatative transversal direction of the ventricular walls to show that the myocardial body contracts actively in a 3-dimensional pattern. This antagonism supports the autoregulation of heart muscle function according to Frank and Starling, preserving ventricular shape, enhances late systolic fast dilation and attenuates systolic constriction of the ventricle wall. Auxotonic dilating forces are particularly sensitive to inotropic medication. Low dose beta-blocker is able to attenuate the antagonistic activity. All myocardial components act against four components of afterload, the hemodynamic, the myostructural, the stromatogenic and the hydraulic component. This complex interplay critically complicates clinical diagnostics. Clinical implications are far-reaching (see Part II, https://doi.org/10.1007/s00059-018-4735-x).

[Antagonistic function of the heart muscle : Part II: Clinical implications]. / Die antagonistische Funktion des Herzmuskels : Teil II: Klinische Auswirkungen.

In the hypertrophic heart the myostructural afterload in the form of endoepicardial networks is predominant, which enhances myocardial hypertrophy. The intrinsic antagonism is derailed. Likewise, the connective tissue scaffold, i.e. the stromatogenic afterload, is enriched in the response to the derailment of antagonism in a hypertrophic heart up to regional captivation of the heart musculature. Due to the selective susceptibility of the auxotonic, contracting oblique transmural myocardial network for low dose negative inotropic medication, this promises to attenuate progress in myocardial hypertrophy. Volume reduction surgery is most effective in reducing wall stress as long as the myocardium is not critically fettered by fibrosis. The use of external mechanical circulatory support is then effective if the heart is supported in its resting mode, which means around a middle width and at minimal amplitude of motion. The takotsubo cardiomyopathy might possibly reflect an isolated, extreme stimulation of the intrinsic antagonism as a response to hormonally induced sensitization of the myocardium to catecholamine. A particular significant conclusion with respect to the diseased heart is that clinical diagnostics need new impulses with a focus on the analysis of local motion patterns and on myocardial stiffness reflecting disease-dependent antagonistic intensity. This would become a relevant diagnostic marker if corresponding (noninvasive) measurement techniques would become available.

Raman micro-spectroscopy analysis of different sperm regions: a species comparison.

STUDY QUESTION: Is Raman micro-spectroscopy a valid approach to assess the biochemical hallmarks of sperm regions (head, midpiece and tail) in four different species? SUMMARY ANSWER: Non-invasive Raman micro-spectroscopy provides spectral patterns enabling the biochemical characterization of the three sperm regions in the four species, revealing however high similarities for each region among species. WHAT IS KNOWN ALREADY: Raman micro-spectroscopy has been described as an innovative method to assess sperm features having the potential to be used as a non-invasive selection tool. However, except for nuclear DNA, the identification and assignment of spectral bands in Raman-profiles to the different sperm regions is scarce and controversial. STUDY DESIGN SIZE, DURATION: Raman spectra from head, midpiece and tail of four different species were obtained. Sperm samples were collected and smeared on microscope slides. Air dried samples were subjected to Raman analysis using previously standardized procedures. PARTICIPANTS/MATERIALS, SETTING, METHODS: Sperm samples from (i) two donors attending the infertility clinic at the Centre of Reproductive Medicine and Andrology; (ii) two C57BL/6 -TgN (ACTbEGFP) 1Osb adult mice; (iii) two adult Cynomolgus monkeys (Macaca fascicularis) and (iv) two sea urchins (Arbacia punctulata) were used to characterize and compare their spectral profiles. Differences and similarities were confirmed by principal component analysis (PCA). MAIN RESULTS AND THE ROLE OF CHANCE: Several novel region-specific peaks were identified. The three regions could be differentiated by distinctive Raman patterns irrespective of the species. However, regardless of the specie, their main spectral pattern remains mostly unchanged. These results were corroborated by the PCA analysis and suggest that the basic constituents of spermatozoa are biochemically similar among species. LIMITATIONS REASONS FOR CAUTION: Further research should be performed in live sperm to validate the detected spectral bands and their use as markers of distinctive regions. WIDER IMPLICATIONS OF THE FINDINGS: Raman peaks that have never been described in the sperm cell were detected. Particularly important are those that are unique to the midpiece as they might be a reference to the identification of sperm mitochondria, whose function is highly correlated with that of sperm. In the future, Raman micro-spectroscopy has the potential to be applied in assessment of male fertility. LARGE SCALE DATA: N/A. STUDY FUNDING AND COMPETING INTEREST(S): This work was supported by BMBF project 'Sperm Ident' (FKZ:13N13024) and the DAAD-CRUP bilateral exchange program (AI A06/16-57213087). S.A. is a recipient of a fellowship from the Portuguese foundation for science and technology (FCT-SFRH/BPD/110160/2015) and R.DC. is a recipient of a DAAD PhD stipend (91590556). There is no competing interest.

Reassembly of adult human testicular cells: can testis cord-like structures be created in vitro?

STUDY QUESTION: Can enzymatically dispersed testicular cells from adult men reassemble into seminiferous cord-like structures in vitro? SUMMARY ANSWER: Adult human testicular somatic cells reassembled into testicular cord-like structures via dynamic interactions of Sertoli and peritubular cells. WHAT IS KNOWN ALREADY: In vitro approaches using dispersed single cell suspensions of human testes to generate seminiferous tubule structures and to initiate their functionality have as yet shown only limited success. STUDY DESIGN, SIZE, DURATION: Testes from 15 adult gender dysphoria patients (mean ± standard deviation age 35 ± 9.3 years) showing spermatogonial arrest became available for this study after sex-reassignment surgery. In vitro primary testicular somatic cell cultures were generated to explore the self-organizing ability of testicular somatic cells to form testis cords over a 2-week period. Morphological phenotype, protein marker expression and temporal dynamics of cell reassembly were analyzed. PARTICIPANTS/MATERIALS, SETTING, METHODS: Cell suspensions obtained by two-step enzymatic digestion were plated onto glass coverslips in 24-well plates. To obtain adherent somatic cells, the supernatant was discarded on Day 2. The culture of the attached cell population was continued. Reassembly into cord-like structures was analyzed daily by microscopic observations. Endpoints were qualitative changes in morphology. Cell types were characterized by phase-contrast microscopy and immunohistochemistry. Dynamics of cord formation were recorded by time-lapse microscopy. MAIN RESULTS AND THE ROLE OF CHANCE: Primary adult human testicular cells underwent sequential morphological changes including compaction and reaggregation resulting in round or elongated cord-like structures. Time-lapse video recordings within the first 4 days of culture revealed highly dynamic processes of migration and coalescence of reaggregated cells. The cellular movements were mediated by peritubular cells. Immunohistochemical analysis showed that both SRY-related high mobility box 9-positive Sertoli and α-smooth muscle actin-positive peritubular myoid cells interacted and contributed to cord-like structure formation. LARGE SCALE DATA: Not applicable. LIMITATIONS, REASONS FOR CAUTION: Owing to scarcity of normal human testicular tissue, testes from gender dysphoria patients were used in the study. The regressed status might influence the experimental responses of primary cells. We observed basic morphological features resembling in vivo testicular cords, however, the proof of functionality (e.g. support of germ cells) will need further studies. WIDER IMPLICATIONS OF THE FINDINGS: The proposed in vitro culture system may open opportunities for examination of testicular cell interactions during testicular tubulogenesis. Further refinement of our approach may enable initiation of ex vivo spermatogenesis. STUDY FUNDING/COMPETING INTERESTS: The work was supported by EU-FP7-PEOPLE-2013-ITN 603568: 'Growsperm'. No conflict of interests is declared.

Pneumatic Distension of Ventricular Mural Architecture Validated Histologically.

Purpose: There are ongoing arguments as to how cardiomyocytes are aggregated together within the ventricular walls. We used pneumatic distension through the coronary arteries to exaggerate the gaps between the aggregated cardiomyocytes, analyzing the pattern revealed using computed tomography, and validating our findings by histology. Methods: We distended 10 porcine hearts, arresting 4 in diastole by infusion of cardioplegic solutions, and 4 in systole by injection of barium chloride. Mural architecture was revealed by computed tomography, measuring also the angulations of the long chains of cardiomyocytes. We prepared the remaining 2 hearts for histology by perfusion with formaldehyde. Results: Increasing pressures of pneumatic distension elongated the ventricular walls, but produced insignificant changes in mural thickness. The distension exaggerated the spaces between the aggregated cardiomyocytes, compartmenting the walls into epicardial, central, and endocardial regions, with a feathered arrangement of transitions between them. Marked variation was noted in the thicknesses of the parts in the different ventricular segments, with no visible anatomical boundaries between them. Measurements of angulations revealed intruding and extruding populations of cardiomyocytes that deviated from a surface-parallel alignment. Scrolling through the stacks of tomographic images revealed marked spiraling of the aggregated cardiomyocytes when traced from base to apex. Conclusion: Our findings call into question the current assumption that cardiomyocytes are uniformly aggregated together in a tangential fashion. There is marked heterogeneity in the architecture of the different ventricular segments, with the aggregated units never extending in a fully transmural fashion. Key Points: • Pneumographic computed tomography reveals an organized structure of the ventricular walls.• Aggregated cardiomyocytes form a structured continuum, with marked regional heterogeneity.• Global ventricular function results from antagonistic forces generated by aggregated cardiomyocytes. Citation Format: • Burg MC, Lunkenheimer P, Niederer P et al. Pneumatic Distension of Ventricular Mural Architecture Validated Histologically. Fortschr Röntgenstr 2016; 188: 1045 - 1053.

UVB irradiation as a tool to assess ROS-induced damage in human spermatozoa.

One of the consequences of oxygen metabolism is the production of reactive oxygen species (ROS) which in a situation of imbalance with antioxidants can damage several biomolecules, compromise cell function and even lead to cellular death. The particularities of the sperm cell make it particularly vulnerable to ROS attack compromising its functionality, mirrored in terms of fertility outcome and making the study of the origin of sperm ROS, as well as the alterations they cause very important. In the present work, we used UVB irradiation, an easy experimental approach known as a potent inducer of ROS formation, to better understand the origin of ROS damage without any confounding effects that usually exist in disease models in which ROS are reported to play a role. To address these issues we evaluated sperm mitochondrial ROS production using the Mitosox Red Probe, mitochondrial membrane potential using the JC-1 probe, lipid peroxidation through BODIPY probe and vitality using PI. We observed that UVB irradiation leads to an increase in sperm mitochondrial ROS production and lipid peroxidation that occur previously to an observable mitochondrial dysfunction. We concluded that sperm UVB irradiation appears to be a good and easily manipulated in vitro model system to study mitochondria-induced oxidative stress in spermatozoa and its consequences, which may be relevant in terms of dissecting the action pathways of many other pathologies, drugs and contaminants, including endocrine disruptors.

Intrathoracic implantation of a continuous flow left ventricular assist device--the microdiagonal blood pump.

AIM: All existing ventricular assist devices are associated with a considerable number of serious complications. We report on our first animal tests with a newly developed microdiagonal blood pump (MDP). METHODS: Six female calves underwent MDP implantation. The inflow and outflow conduit were anastomosed to the left atrium and the descending aorta. The MDP was placed in the left phrenicocostal sinus. Pump flow was adjusted to 2-3 L/min. Hemodynamic and echocardiographic data as well as blood samples were measured over the test period of 7 days. Thereafter, all internal organs and the pump were explanted for thorough examination. RESULTS: Mean arterial (90.5+/-12.1102.7+/-8.7 mmHg) and mean pulmonary arterial (17.3+/-2.420.6+/-2.7 mmHg) pressures remained stable during the test period. Cardiac output (5.4+/-0.5 = or >3.5+/-0.5 L/min) decreased postoperatively due to partial unloading of the heart. Left ventricular end-diastolic (4.4+/-0.5= or >3.8+/-0.4 cm) and end-systolic (3.4+/-0.5 =or >2.9+/-0.4 cm) diameters decreased after MDP implantation and did not change during the test period. Mean number of platelets (550+/-57 =or >350+/-86 x 103/microL) and hemoglobin (13.2+/-1.3 = or >11.9+/-0.8 g/dL) decreased perioperatively due to surgical reasons and recovered in the postoperative course. Free hemoglobin was slightly enhanced in the postoperative course. CONCLUSION: Our results demonstrate that the MDP is suitable for intrathoracic implantation and provides a reliable left ventricular unloading.

The forces generated within the musculature of the left ventricular wall.

OBJECTIVES: To test the hypothesis that two populations of myocardial fibres-fibres aligned parallel to the surfaces of the wall and an additional population of fibres that extend obliquely through the wall-when working in concert produce a dualistic, self stabilising arrangement. METHODS: Assessment of tensile forces in the walls of seven porcine hearts by using needle probes. Ventricular diameter was measured with microsonometry and the intracavitary pressure through a fluid filled catheter. Positive inotropism was induced by dopamine, and negative inotropism by thiopental. The preload was raised by volume load and lowered by withdrawal of blood. Afterload was increased by inflation of a balloon in the aortic root. The anatomical orientation of the fibres was established subsequently in histological sections. RESULTS: The forces in the fibres parallel to the surface decreased 20-35% during systolic shrinkage of the ventricle, during negative inotropism, and during ventricular unloading. They increased 10-30% on positive inotropic stimulation and with augmentation in preload and afterload. The forces in the oblique transmural fibres increased 8-65% during systole, on positive inotropic medication, with an increase in afterload and during ventricular shrinkage, and decreased 36% on negative inotropic medication. There was a delay of up to 147 ms in the drop in activity during relaxation in the oblique transmural fibres. CONCLUSION: Although the two populations of myocardial fibres are densely interwoven, it is possible to distinguish their functions with force probes. The delayed drop in force during relaxation in obliquely oriented fibres indicates that they are hindered in their shortening to an extent that parallels any increase in mural thickness. The transmural fibres, therefore, contribute to stiffening of the ventricular wall and hence to confining ventricular compliance.

A critical evaluation of results of partial left ventriculectomy.

BACKGROUND: Because of the variation in the surgical procedures designed to reduce ventricular radius, along with differences in hospital care, it is difficult to disentangle the factors that may contribute to the success or failure of the partial left ventriculectomy. METHODS AND RESULTS: We undertook partial left ventriculectomy in 18 patients, 10 suffering from idiopathic dilated cardiomyopathy and 8 from ischemic heart disease. We assessed the amount of reduction in wall stress, the systolic thickening of the ventricular wall, and the extent of connective tissue in the excised segment of the wall. Of the overall group, six patients died, three from infarction, one of stroke, one with asystole, and one with ventricular fibrillation. The mean decrease in measured mesh tension was 40% (p < 0.001). Most patients exhibited improvements postoperatively in terms of the systolic thickening of the posterior and superior free walls of the left ventricle. In those in whom the events could be monitored, life-threatening arrhythmias posed complications in three of four patients with ischemic heart disease, and in two of six patients suffering from idiopathic dilated cardiomyopathy. In one patient, death was associated with a transmural alignment of fibrous tissue. CONCLUSIONS: Our measured reductions in myocardial mesh tension were in keeping with the anticipated theoretical reduction in wall stress expected from partial ventriculectomy. The basic concept underscoring surgical maneuvers to reduce ventricular radius, therefore, is sound. A potential trap is the resection of the marginal artery. Critical myofibrosis was a rare complication. Arrhythmias, which are common, can successfully be treated by implantation of antitachycardic and defibrillatory devices.

Does a suitable animal model for research on partial left ventriculectomy exist? Animal models for PLV.

Partial left ventriculectomy is a new surgical option quickly introduced into clinical use worldwide for treating end-stage heart failure in patients with dilated cardiomyopathy. Due to the overwhelming success of this new kind of surgical treatment for dilated cardiomyopathy, experimental research on the physiological and pathophysiological basis was initially not performed. Now, demands for an appropriate animal model have arisen more and more since the outcome of patients treated by partial left ventriculectomy has differed considerably. This review summarizes available experimental models for heart failure in large animals, and discusses their suitability for research on partial left ventriculectomy.

Treatment strategy and perioperative risk in patients with idiopathic thrombocytopenic purpura undergoing cardiac surgery.

Patients suffering from idiopathic thrombocytopenic purpura undergoing cardiac surgery with extracorporeal circulation may be at increased risk for postoperative bleeding complications. Therefore, different treatment strategies, such as conservative treatment, splenectomy, transfusion of platelet concentrates and therapy with immunoglobulins have been proposed. We report on our experiences with preoperative immunoglobulin treatment in selected patients with special regard on the optimal dosage.

Late ventricular structure after partial left ventriculectomy.

Nine months after partial ventriculectomy, a 53-year-old man died of progressive heart failure. His heart was examined to determine the alignment of the muscle fibers around the ventricular scar, which was 11 cm long, 1.3 cm thick and 4 cm wide. The scar reached 2 to 12 mm beyond the surgical suture line. The fibers in the middle and subendocardial layers were malaligned, resulting in convergence, compression and regional necrosis.

Preoperative immunoglobulin treatment in patients with Werlhof's disease undergoing cardiac operation.

BACKGROUND: Patients with Werlhof's disease and undergoing a cardiac surgical procedure with cardiopulmonary bypass are at increased risk for bleeding complications. We report the usefulness of preoperative immunoglobulin treatment in selected patients. METHODS: Between May 1995 and July 1998, 10 patients with Werlhof's disease underwent a cardiac surgical procedure with cardiopulmonary bypass in our department. Five patients with mean platelet counts of less than 80x10(9)/L received immunoglobulin therapy preoperatively (group 1). The other 5 patients with mean platelet counts higher than 80x10(9)/L were not so treated (group 2). RESULTS: In group 1, mean platelet count increased from 54x10(9)/L 5 days before operation to 112x10(9)/L after immunoglobulin treatment (p = 0.018) and did not fall to less than 60x10(9)/L postoperatively. Patients in group 1 received 16 units of packed red blood cells and 5 units of platelet concentrate. Patients in group 2 required 24 units of packed red blood cells, 5 units of platelet concentrate, and 23 units of fresh frozen plasma. Only 1 patient (group 2) had a surgical bleeding complication that required reexploration. Mean drainage loss was 1,100 mL in group 1 and 1,210 mL in group 2. CONCLUSIONS: Our data demonstrate that immunoglobulin treatment of patients with Werlhof's disease and mean platelet counts of less than 80x10(9)/L significantly augments platelet counts preoperatively. It may be useful in selected patients.

Immediate effects of partial left ventriculectomy on left ventricular function.

BACKGROUND: Attempts to prolong life or to improve the quality of life by partial left ventriculectomy in patients suffering from dilated cardiomyopathy have yielded strikingly variable results in leading surgical centers. HYPOTHESIS: The outcome of patients after partial left ventriculectomy depends on intraoperative myocardial protection together with appropriate long-term pharmacotherapy. We further assume that partial removal of the fibrotic ventricular wall may lead to a particularly inhomogeneous pattern of wall stress, giving rise to the potential of a paradoxical increase in wall stress and the creation of arrhythmogenic foci. METHODS: During surgery in 24 patients, local mesh tension was measured using needle-force probes in up to five sites within the left ventricular wall before and after resection of the interpapillary mural segment. The data were used to calculate regional peak developed force and to identify any differences in the timing of local mechanical activity between the measured regions. RESULTS: Mean decrease in regional wall stress was 42% (76 sites of measurement). However, we discovered a paradoxical increase of 42% in 18 sites of measurement. The time delay in the onset of force development between the measured regions prior to surgery was 0 msec in 10 patients, up to 30 msec in 7 patients, and beyond 80 msec in 7 patients. After resection, the time delay increased considerably in incidence and duration. CONCLUSION: Ventriculectomy is an effective means of reducing wall stress. The unexpectedly high incidence of inhomogeneities in wall stress after asymmetrical surgical ventricular remodeling, currently typical for the classical Batista procedure, together with the asynchronous regional ventricular function that we found to increase after partial left ventriculectomy, needs further elucidation by electrophysiological investigations.

A silicone rubber mould of the heart.

The macroscopical structure of the ventricular myocardium has been an unsolved problem since the XVIth century, when Anatomy started as an authentic science. Since then the spatial organization of the myocardial fibres has represented, as Pettigrew says, "an arrangement so unusual and perplexing, that it has long been considered as forming a kind of Gordian knot in Anatomy. Of the complexity of the arrangement I need not speak further than to say that Vesalius, Albinus, Haller and De Blainville, all confessed their-inability to unravel it". What is shown in the present paper is the result of an anatomical work, developed over 43 years, by means of which it has been shown that the ventricular myocardial mass consists of a band, curled in a helical way, which extends from the pulmonary artery to the aorta. This is illustrated by a silicone rubber model cast from an actual unrolled myocardial band.

The heart muscle's putative "secondary structure'. Functional implications of a band-like anisotropy.

Opinions are divided as to whether the rope-like secondary structure, which Torrent-Guasp dissected out of the myocardial body by the blunt unwinding technique (BUT) reveals some kind of functional compartmentation of the heart muscle. The myocardial fibres are aligned parallel to the fibre disruption (cleavage) plane, along which the band has been prepared but they are not necessarily aligned parallel to the long axis of the band. Inconsistencies in the myocardial rope model arise from the obligatory zones of transmural inflection, which are obvious in the base and the apex of both ventricles. They are, however, merely discernible in the midzone of the left ventricular cone. The investigator experienced in BUT knows that the cleavage plane is not unique. We doubt the assumption that the rope structure is the predominant stress transmission pathway, because the fibre strand peel-off technique (SPOT) delivers irregular fibre disruption planes which are definitely different from those which Torrent-Guasp prepares. The rope-like fibre arrangement could be just a redundant structure, a remnant of past developmental steps without, however, any functional implication to the human heart. On the other hand, peeling-off fibre strands from the ventricular wall produces deeply perforating, i.e., oblique transmurally grooved surfaces. Putative functions of force transmission in an oblique transmural direction are (1) ventricular dilation as a function of the variable inclination angle with respect to the epicardial surface, (2) monitoring of ventricular wall stress and ventricular size and (3) segmental stiffening which could serve other dependent segments as a punctum fixum.

The heart's fibre alignment assessed by comparing two digitizing systems. Methodological investigation into the inclination angle towards wall thickness.

Myocardial contractile pathways which are not aligned strictly parallel to the heart's epicardial surface, give rise to forces which also act in the ventricular dilating direction. We developed a method which allows us to assess any fibre orientation in the three-dimensional myocardial weave. Decollagenized hearts were prepared by peeling-off fibre strands, following their main fibre orientation down to near the endocardium. In the subepicardium the strands followed a course more or less parallel to the epicardium, whereas from the mid-wall on they tended to dive progressively deeper into the wall. The preparation displays more or less rugged surfaces rather than smooth layers. The grooves and crests on the exposed surfaces were sequentially digitized by two methods: (1) Using a magnet tablet (3 Draw Digitizer System, Polhemus, Cochester VTO 5446, USA) on a dilated pig heart we manually followed the crests using a stylus, handling each groove and crest as an individual contractile pathway. (2) A constricted cow heart was digitized using a contact-free optical system (opto TOP, Dr. Breuckmann, Meersburg, Germany), which is based on the principle of imaging triangulation. Using specially developed software the inclination angles of selected crests and grooves with respect to the epicardial surface were calculated. The two digitizing methods yield comparable results. We found a depth- and side-specific weave component inclined to the epi-endocardial direction. This oblique netting component was more pronounced in the inner 1/3 of the wall than in the subepicardium. The inclination angle probably increases with increasing wall thickness during the ejection period. Manual digitizing is an easy and fast method which delivers consistent results comparable with those obtained by the cumbersome high resolution optical method. The rationales for the assessment of transmural fibre inclination are (1) the putative existence of dilating forces inherent in the myocardial weave and (2) the possible overproportional increase in the oblique transmural weave component during myocardial hypertrophy, which would entail a reduction in efficiency of ventricular performance in terms of haemodynamic work.