[Clinical pathology in the Hanseatic City of Lübeck from the planning of an institute to the formation of the "Medizinische Hochschule"]. / Die klinische Pathologie in der Hansestadt Lübeck. Von der Planung des Instituts bis zur Gründung der Medizinischen Hochschule.

The formation of an university institute of pathology at the Hanseatic city of Lübeck preceded a long, changeful planning, accompanied by a chain of historical events. In the 19th century, Lübeck owned seven different places for clinical or forensic autopsies. Thus, in the first years of the 20th century it was generally called for the formation of a "Central Institute of Pathology" that was completed after all in the year 1927. Two years later, Eberhard von Praun became head of the institute and was immediately confronted with the "Lübeck-disaster". He was followed 1935 by Ernst Jeckeln, who identified the "Enteritis necroticans" which appeared in North Germany in the first years after the "Second World War". He called the disease "Darmbrand", a term that is since then internationally used. Jeckeln became the first "Ordinarius for Pathology" at the "Medical Academy Lübeck", which was associated to the "University of Kiel". His long-time colleague was Alfred Wegener, who identified the "Wegener's granulomatosis". In the year 1972 Alfred Gropp became head of the institute, an all-round scientist and pathologist, e.g. well-known in the field of developmental pathology. During his term, the "Medical Academy" got the university status and was named "Medizinische Hochschule Lübeck".

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.

[Transmission electron microscopy image of wear particles of joint endoprostheses and ultrastructural cell changes]. / Transmissionselektronenmikroskopische Darstellung der Abriebpartikel von Gelenkendoprothesen und von ultrastrukturellen Zellveränderungen.

Wear particles from joint endoprostheses vary considerably in size, and may be detectable in tissue only by electron microscopy. Wear debris plays a central role in the non-infectious late loosening of prostheses, and it has been estimated that the submicron particles induce increased liberation of mediators of osteolysis by activated macrophages. From the types of prostheses currently in use, bone cement and polyethylene particles greatly predominate over metallic and ceramic particles. Since it had formerly not been possible to reliably identify wear particles in the transmission electron microscopy, and descriptions of them in the literature varied considerably, we analysed ultrathin sections obtained from periprosthetic tissue containing wear particles previously identified by laser microprobe mass analysis. Using this method, it proved possible to classify almost all the wear particles detected in the electron microscope, to determine their size range and to represent the cellular alterations caused by them.

[Severe 46,XY virilization deficit due to 17beta-hydroxysteroid dehydrogenase deficiency]. / Schwere 46,XY Virilisierungsstörung durch 17beta-Hydroxysteroid Dehydrogenase 3 Mangel.

BACKGROUND: 17beta hydroxysteroid dehydrogenase deficiency is a presumable rare cause for a severe virilization disorder in children with 46,XY karyotype due to a defect in the testicular testosterone biosynthesis from androstenedione. PATIENT: We report on a 14 year old child with 46,XY karyotype with a predominantly female phenotype. RESULTS: Hormonal analysis showed low values for androstenedione and testosterone before and after stimulation with human chorionic gonadotropin, however, the androstenedione/testosterone ratio was elevated. Molecular genetic analysis proved the diagnosis of 17beta-hydroxysteroid dehydrogenase deficiency due to a homozygous mutation (325+4 A-T) in the HSD17B3-gene, which leads to an aberrant splicing process. CONCLUSIONS: This case demonstrates that in addition to a meticulous steroid analysis a direct molecular genetic analysis can be helpful in the diagnosis of 17beta-hydroxysteroid dehydrogenase deficiency.

Increased frequency of X-bearing sperm in males from an infertility clinic: analysis by two-color fluorescence in situ hybridization.

Semen samples from 34 men visiting the Lübeck infertility clinic were investigated using a two-color FISH method to determine the ratio of X- and Y-bearing sperm. The overall ratio was significantly shifted to a preponderance of X-containing sperm. A statistical comparison with seven reports from the literature which included 53 normal probands demonstrated in our patients a significant tendency of a preponderance of X-bearing sperm and significantly less Y-bearing sperm. Furthermore, the Lübeck sperm samples are remarkably more heterogeneous in respect to their variability of X- and Y-bearing spermatozoa than in the other mentioned studies with normal probands. These phenomena have to be evaluated in further studies on groups of infertile males showing similar infertility histories.

Primary epithelioid angiosarcoma of the adrenal gland case report and review of the literature.

Primary angiosarcoma of the adrenal gland is extremely rare. Here, we report on a 70-year-old man with an angiosarcoma of the right adrenal gland who died 3 weeks after tumor resection due to intestinal infarction and acute renal failure. No metastases were found at autopsy. Histologically, the tumor showed a predominantly epithelioid differentiation. Immunohistochemical examination revealed positive reactivity for cytokeratin, epithelial membrane antigen, vimentin, factor VIII-related antigen, CD31, CD34 and Ulex europaeus agglutinin-I. Features of endothelial origin were also demonstrable by electron microscopy. The differential diagnosis of this uncommon neoplasm is discussed. The present case emphasizes problems in differential diagnosis that arise from its epithelioid differentiation. A review of the literature underlines the poor clinical outcome of adrenal angiosarcoma.

[Identification of wear particles of joint prostheses in tissues using laser microprobe mass analysis (LAMMA)]. / Identifizierung von Abriebpartikeln der Gelenkendoprothesen im Gewebe mit Hilfe der Laser-Mikrosonden-Massenanalyse (LAMMA).

The definite identification of wear particles from joint prostheses is of great importance for the development of joint replacement, as the type and quantity of different wear particles gives information on the wear resistance of implant materials. From the types of prostheses nowadays in use polyethylene wear of the sockets, bone cement wear, metallic and ceramic wear can be generated. Whereas polyethylene wear can be easily identified by its bright luminescence in polarized light and its characteristic configuration, the distinction of the small granular wear particles of the bone cement, metal and ceramic by light microscope is difficult. The laser microprobe mass analysis (LAMMA) is a method, which allows the analysis of single light microscopically detectable wear particles in tissues. Not only contrast medium particles of the bone cements (zirconium oxide or barium sulfate) but also metallic and aluminum oxide particles could be definitely identified within the pseudocapsules as well as in regional lymph nodes by LAMMA-analysis, whereby the bone cement wear predominated. In addition, the distinction between organic substances (as blood degradation products), which may appear similar to wear particles in configuration and colour, and the foreign material is also possible with this method.

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.

Use of frozen-thawed testicular sperm for intracytoplasmic sperm injection.

OBJECTIVE: To determine the feasibility of using frozen-thawed testicular spermatozoa for intracytoplasmic sperm injection. DESIGN: Prospective clinical study. SETTING: A university hospital. PATIENT(S): One hundred seventy-five azoospermic men participating in a routine intracytoplasmic sperm injection program. INTERVENTION(S): The men underwent testicular biopsy for cryopreservation of tissue to be used in consecutive intracytoplasmic sperm injection treatment cycles. Their female partners underwent controlled ovarian hyperstimulation for conventional IVF treatment. MAIN OUTCOME MEASURE(S): Fertilization and pregnancy rates. RESULT(S): In 77% of the patients, spermatozoa could be harvested from the testis by an open testicular biopsy technique and used for intracytoplasmic sperm injection after freezing and thawing of testicular tissue. Histopathologic evaluation revealed a Sertoli cell-only pattern in 21%, maturation arrest in 60%, and hypospermatogenesis in 19% of the patients. In 2. 9% of the patients, carcinoma in situ or a germ cell tumor was detected. In all patients, viable spermatozoa could be visualized after the tissue samples were thawed. One hundred thirty-five intracytoplasmic sperm injection treatment cycles were performed, with a fertilization rate of 45% and a clinical pregnancy rate of 30% per oocyte retrieved. CONCLUSION(S): The use of frozen-thawed testicular tissue allows ovarian stimulation of the female partner to be timed and avoids cancellation of ovum pick-up when spermatozoa cannot be retrieved.

[Malakoplakia of the colon]. / Malakoplakie des Kolons.

We report on a 62-year-old female patient with melena in which polypoid lesions of the cecum were discovered endoscopically. Histological examination of mucosal biopsies revealed an inflammatory process with lots of histiocytes and so-called Michaelis-Gutmann bodies, leading to the diagnosis of a malakoplakia of the colon. No other organs were found affected. In the course of an antibiotic therapy, there was no melena detectable over a period of six months. Malakoplakia, an inflammation usually affecting the urogenital tract, is rarely found in the colon, with only 35 cases published until now. It is frequently associated with other diseases like neoplastic or inflammatory disorders, immune defect syndromes or heroin abuse. This spectrum is expanded by our report in which a long-standing alcohol abuse was found as an attendant disease.

Intracytoplasmic injection of round and elongated spermatids from azoospermic patients: results and review.

Microinjection is established as the method of choice in the treatment of severe male factor infertility as well as in azoospermic patients. Recent studies have shown that fertilization and cleavage can be achieved by injection of ejaculated as well as testicular elongated spermatids into oocytes. Here we report on the two first pregnancies worldwide resulting from elongated spermatid injection from frozen-thawed testicular tissue. Four patients with complete Sertoli cell-only syndrome (SCOS) and two with spermatogenetic maturation arrest were included in our microinjection programme. Tissues from open testicular biopsies were cryopreserved until the time of follicle puncture. A total of 67 oocytes were harvested. In the two patients with maturation arrest, cryopreserved elongated spermatids were successfully injected, while in two of the other four SCOS patients only cryopreserved round spermatids were available to be injected into the oocytes. Out of 18 injected oocytes, 10 were fertilized in the first group, while nine out of 49 injected oocytes showed fertilization and cleavage in the second group. Two clinical pregnancies were achieved with elongated spermatids from frozen-thawed testicular tissue, while no pregnancy was established in the case of round spermatids. This study confirms that fertilization, cleavage and pregnancy can be successfully achieved in cases with spermatogenetic maturation arrest by injecting cryopreserved elongated spermatids into oocytes. The literature on pregnancies following spermatid injection, as well as the problems using this technique and possible risks, are discussed.

Pachytene chromosomes in trisomy 19 male mice with Robertsonian translocations.

Three male mice with trisomy 19 induced by a Robertsonian translocation system were used for the study of meiotic prophase cells and germ cell differentiation. Present in these males were two Robertsonian chromosomes each with a chromosome 19 arm in common, two acrocentric chromosomes corresponding to the second arms of the two Rbs and one acrocentric chromosome 19. These five chromosomes showed a wide range of meiotic pairing configurations. One particular observation was the formation of a true double synaptonemal complex (SC) with three lateral axes and two central elements, which joined the three chromosomes 19 together. Integration of the acrocentric chromosome 19 in a complex pentavalent configuration was seen in 45% of the pachytene nuclei. The proportion of spermatocytes showing association between a quadrivalent and the acrocentric no. 19 was 26%. In 29% of the nuclei, the acrocentric no. 19 was free, integrated or associated with the XY complex, paired with the X chromosome or associated with a bivalent. Finally, in 57% of pachytene cells, the meiotic multivalents or the free univalent 19 were associated with the proximal part of the X chromosome or integrated in the sex chromatin. Therefore, the question arises with regard to the fate of these spermatocytes. The testicular histology shows an arrest of germ cell development at the spermatocyte stage. Several mechanisms seem to be the cause of germ cell depletion in a sequence of different, impaired developmental processes.

Thanatophoric dysplasia type II: new entity?

There is some question about whether the two forms of thanatophoric dysplasia (TD), Type II with and Type I without cloverleaf skull, belong to the same entity. Thus, we investigated one 6-day-old TD with cloverleaf skull using examination of the external phenotype, radiology, autopsy, skeleton preparation, large section histology, detailed section histology, and ultrastructure. The loss of the three-phase contours-characteristics for Type I (54)--in certain metaphyses, the absence of the perichondral spurs to some extent, and their substitution by a structure similar to the perichondral "ring of Lacroix" have a suggested origin in normal cartilage-bone tissue. The same mechanism is postulated (a) for the appearance of less bent or normally shaped tubular bones compared with TD Type I, and their corresponding increased mechanical stability, and (b) for the less amount of platyspondyly in Type II than in Type I. We suggest that the malformation of the cloverleaf skull has its origin in the promontory growth of the relatively normal cartilage-bone tissues at the skull base resulting in an early synostosis and a consecutive fusion of the cranial sutures. The ultrastructural analysis of chondrocytes demonstrates the significant contribution of electron microscopy for TD studies. We suggest that pathologically altered light chondrocytes accounts for plump cross-striated collagen fibrils, the reduced cellular proliferation, and the impaired formation of columnar and hypertrophic zones. It is clear that normal cartilage-bone tissue distributed among "thanatophoric" tissue is the reason for the differences between Type I and Type II. Hypotheses are presented that explain this tissue mosaicism. Thus, TD Type I and TD Type II do not represent two different entities but the same entity with varying features due to mutational events occurring at different times.

Fine needle tissue aspiration for accurate localization of histological specimens from complex structures.

A procedure is presented for exact, detailed comparison of light and electron microscopic analyses of tissues with complex architecture. Earlier techniques require one to make drawings of tissue pieces to be analyzed by electron microscopy to permit rough localization of the origin of the tissue pieces. Specifically, exact analysis of fetal cartilage and bone is hampered by the complicated arrangement of both tissue components, severely limiting the assessment of electron microscopic analyses. The advantage of the technique described here is that it allows precise localization of the tissue sample in the original tissue area. Punches 1 min in diameter were obtained from femora and coxae with a syringe and embedded for light and electron microscopy. The remaining tissue with its exactly defined punctures is prepared for standard histology. Human fetal cartilage and bone tissue were used to demonstrate this technique, but this procedure may be used for other kinds of tissues.

Cyclophosphamide-induced aberrations of chromosome pairing in pachytene oocytes.

The aim of this study was to characterise damage to synaptonemal complexes in oocytes following cyclophosphamide exposure. Pregnant mice were treated with three different doses of cyclophosphamide (10, 30 and 50 mg/kg body weight) at day 13 of gestation, when oogonia and very early meiotic cells in the female fetuses are found. Primary oocytes were analysed by light- and electron microscopy at gestational day 17 to reveal effects of the alkylating agent on the chromosomal pairing behaviour. Our pachytene analysis demonstrated that the fraction of cells with lesions of synaptonemal complexes, partial asynapsis and desynaptic bivalents were significantly increased over the levels in the control group. The frequency of alterations was similar at doses of 30 and 50 mg/kg. In addition, a significant increase in frequency of univalents over the base level became evident, showing the highest incidence on a dose of 50 mg/kg.

Scoliosis and congenital anomalies associated with Klippel-Feil syndrome types I-III.

STUDY DESIGN: This investigation was aimed at characterizing anomalies and syndromes associated with Klippel-Feil syndrome in a large group of patients. The authors evaluated the clinical and radiographic features, documented the associated anomalies, and registered the type of treatment. OBJECTIVE: The anomalies or syndromes and the development of scoliosis were correlated to the type of Klippel-Feil syndrome. MATERIAL AND METHODS: In a cross-sectional study, the authors reviewed data from 57 patients with Klippel-Feil syndrome treated over 25 years at the Department for Orthopedics of the University of Heidelberg. The patients (17 males and 40 females; average age of the first contact, 12 years) were classified into three types according to the description of Feil in 1919. RESULTS: Klippel-Feil syndrome Type I (fusion of cervical and upper thoracic vertebra with synostosis) and Type II (isolated cervical spine) corresponded to 40% and 47% of patients, respectively. Type III (cervical vertebra associated with lower thoracic or upper lumbar fusion) was displayed in 13% of the patients only. The authors found a variety of combinations of Klippel-Feil syndrome and other anomalies in the patients examined in this study, with 67% of the patients characterized by an association with other disorders or syndromes. Of the patients, 70% showed scoliosis. Its degree depended on the type of Klippel-Feil syndrome. Scoliosis in Type I correlated with 31 degrees (Cobb angle), in Type III with 23 degrees, and in Type II with 9 degrees only. Thus, Type II, with isolated cervical fusion, shows a low risk for scoliosis. CONCLUSION: This study increases knowledge of a wide range of anomalies and syndromes identified in association with Klippel-Feil syndrome. A special finding of the study was a correlation between the degree of scoliosis and Klippel-Feil syndrome Types I, II, and III.

[A new punch technique of cartilage-bone tissue for morphological studies in orthopedics]. / Eine neue Stanzentechnik am Knorpel-Knochengewebe für morphologische Untersuchungen in der Orthopädie.

A new technique is presented for the analysis of tissues which allows an exact, detailed comparison of light and electron microscopy. For this purpose, the previous techniques required the cutting of tissue pieces from the original tissue compound and the preparation of drawings as a rough localisation of the tissue pieces removed for analysis. Due to the processes of cutting and drawing, only an approximate localisation of the tissue investigated by electron microscopy was practicable. Specifically, the exact analysis of fetal cartilage-bone-tissue was mostly not possible due to the complicate arrangement of both tissue components. Therefore, the assessment of electron microscopic analyses was severely limited. The new technique, presented in this study, is characterised by a removal of punches 1 mm in diameter from the original tissue for electron microscopy. The remaining tissue with its exact defined punch-punctures is prepared for standard histology. Thus, the specific advantage of this new method is the precise attribution of the punched tissue to the original tissue group. Human fetal cartilage-bone-tissue has been used to demonstrate this punch-technique and the specific light- and electron microscopic preparations.

Synaptonemal complex damage in fetal mouse oocytes induced by ionizing irradiation.

Fetal female mice were exposed to ionizing irradiation of 2 Gy in a single dose at days 14, 16, and 17 of gestation. Synaptonemal complexes of primary oocytes were analyzed on day 17. It has been demonstrated that electron beam irradiation of early oocytes on day 14 with 2 Gy is accompanied by a duplication of atretic cells. A significant increase in fragmentations of the synaptonemal complexes over the base level became evident when mice were exposed to irradiation on days 16 and 17 of gestation. Frequencies of multivalent formation and univalents were not increased over the levels in the control group. Reduction of fertility and malsegregation of chromosomes may be a reflection of the consequences of the observed nuclear lesions.

Synaptonemal complexes of chains and rings in mice heterozygous for multiple Robertsonian translocations.

Complex Robertsonian translocation heterozygosities in the mouse have been used to test different hypotheses regarding the correlation between male hybrid sterility and chromosomal abnormality. Synaptonemal complexes of meiotic super-chains and super-rings involving 15 to 18 metacentric chromosomes were studied in relation to spermatogenic histology. Both types of multivalents showed a characteristic pachytene pattern of alternating paired and non-paired segments. The amount of unpaired segments in rings was about 18% and in chains about 23% of the total length of multivalent chromosomes. The meiotic chains were associated with the proximal part of the X chromosomes in more than 60% of pachytene cells; a similar tight proximity of rings with X or Y chromosomes was never found. Complete arrest of germ cell maturation correlated with super-chains and inconspicuous testicular histology with super-rings. This demonstrates that an excessive amount of unpaired chromosomal axes does not lead per se to male infertility through gametogenic breakdown. On the contrary, the results clearly indicate spermatogenic impairment in this system of multimetacentric heterozygosity as a reflection of X chromosome super-chain interference.