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The existence of an amniotic fluid microbiota (i.e., a viable microbial community) in mammals is controversial. Its existence would require a fundamental reconsideration of fetal in utero exposure to and colonization by microorganisms and the role of intra-amniotic microorganisms in fetal immune development as well as in pregnancy outcomes. In this study, we determined whether the amniotic fluid of mice harbors a microbiota in late gestation. The profiles of the amniotic fluids of pups located proximally or distally to the cervix were characterized through quantitative real-time PCR, 16S rRNA gene sequencing, and culture (N = 21 dams). These profiles were compared to those of technical controls for bacterial and DNA contamination. The load of 16S rRNA genes in the amniotic fluid exceeded that in controls. Additionally, the 16S rRNA gene profiles of the amniotic fluid differed from those of controls, with Corynebacterium tuberculostearicum being differentially more abundant in amniotic fluid profiles; however, this bacterium was not cultured from amniotic fluid. Of the 42 attempted bacterial cultures of amniotic fluids, only one yielded bacterial growth – Lactobacillus murinus. The 16S rRNA gene of this common murine-associated bacterium was not detected in any amniotic fluid sample, suggesting it did not originate from the amniotic fluid. No differences in the 16S rRNA gene load, 16S rRNA gene profile, or bacterial culture were observed between the amniotic fluids located Proximally and distally to the cervix. Collectively, these data indicate that, although there is a modest DNA signal of bacteria in murine amniotic fluid, there is no evidence that this signal represents a viable microbiota. While this means that amniotic fluid is not a source of microorganisms for in utero colonization in mice, it may nevertheless contribute to fetal exposure to microbial components. The developmental consequences of this observation warrant further investigation.

The existence of an amniotic fluid microbiota (i.e., a viable microbial community) in mammals is controversial. Its existence would require a fundamental reconsideration of the role of intra-amniotic microbes in fetal development and pregnancy outcomes. In this study, we determined whether the amniotic fluid of mice harbors a microbiota in late gestation. Bacterial profiles of amniotic fluids located proximally or distally to the cervix were characterized through quantitative real-time PCR, 16S rRNA gene sequencing, and culture (N = 21 mice). These profiles were compared to those of technical controls for background DNA contamination. The load of 16S rDNA in the amniotic fluid exceeded that in controls. Additionally, the 16S rDNA profiles of the amniotic fluid differed from those of controls, withCorynebacterium tuberculostearicumbeing differentially more abundant in amniotic fluid profiles; however, this bacterium was not cultured. Of the 42 total bacterial cultures of amniotic fluids, only one yielded bacterial growth –Lactobacillus murinus. The 16S rRNA gene of this common murine-associated bacterium was not detected in any amniotic fluid sample, suggesting it did not originate from the amniotic fluid. No differences in 16S rDNA load, 16S rDNA profile, or bacterial culture were observed between amniotic fluids located proximal and distal to the cervix. Collectively, these data show that, although there is a modest DNA signal of bacteria in murine amniotic fluid, there is no evidence that this signal represents a viable microbiota. These findings refute the proposed role of amniotic fluid as a source of microorganisms forin uterocolonization.IMPORTANCEThe prevailing paradigm in obstetrics has been the sterile womb hypothesis, which posits that fetuses are first colonized by microorganisms during labor and/or the vaginal delivery process. However, it has been suggested that fetuses are consistently colonizedin utero. One proposed source of colonizers is the amniotic fluid surrounding the fetus. This concept has been derived primarily from investigations that relied on DNA sequencing. Due to the low microbial biomass of amniotic fluid, such studies are susceptible to influences of background DNA contamination. Additionally, even if there is a microbial DNA signature in amniotic fluid, this is not necessarily reflective of a resident microbiota that could colonize the mammalian fetus. In the current study, using multiple microbiologic approaches and incorporating technical controls for DNA contamination, we show that, although there is a low abundance bacterial DNA signal in amniotic fluid, this does not translate to the presence of viable bacteria.

Background: According to the ROTAS study most of the improvement in visual acuity (VA) during amblyopia therapy of children aged 3 to 8 years occurs during the first 6 to 8 weeks 1. Sattler reported a VA gain in 11-year olds even during the second year of treatment 2. So far there are no standards concerning the intensity and duration of the treatment of patients older than 7 years of age. After a report on electronic monitoring of occlusion treatment in patients aged 7 to 16 years for 4 months 3, we now analyse whether this age group benefits from a longer-lasting treatment. Materials and Methods: In this pilot study the progression of VA was analysed in 11 patients (age range 7.18 to 15.76 years; median 11.42 years) during 12 months of occlusion therapy (types of amblyopia: 5 anisometropic, 1 strabismic, 5 combined). The daily occlusion times were recorded using the occlusion dose monitor (ODM) 4. At the beginning of treatment the prescription of the occlusion regime (median) was 6 h/d (range 4 to 7 h/d), the (decimal) VA 0.2 (range 0.02 to 0.63) for single and 0.16 (range 0.02 to 0.8) for crowded optotypes. Results: The recorded occlusion time (median) was 4.4 h/d during the 12 months of treatment, the VA gain (median) was 0.4 log units for single (range 0.2 to 0.7 log units) and 0.3 for crowded optotypes (range 0.1 to 0.6). During the period of 4 to 12 months of treatment (received occlusion 4.12 h/d) the VA gain was 0.1 log units for single and for crowded optotypes. The maximum VA gain during the interval of 4 to 12 months of treatment was 0.2 log units, both single and crowded. The interocular difference for crowded VA (median) decreased from 0.9 to 0.6 log units during treatment, however only one patient achieved an interocular difference of < 0.2 log units. Conclusion: The patients presented here were able to integrate daily occlusion lasting several hours and the electronic monitoring of occlusion treatment into their daily routine over a period of 12 months. During this period the VA of all included types of amblyopia improved significantly, both from a clinical and statistical point of view. Further long-term studies using ODMs with larger groups of patients may identify factors for success of treatment, reveal the long-term stability of the improvement and contribute to a standardised treatment in this age group.

Lung resection showing giant cell (African) histoplasmosis was performed because of suspected lung tumour in a patient who had lived mainly in West Africa. A primary focus or signs of localisation in other organs were suspected radiographically in the maxilla and in the skeleton. However, they could not be confirmed as the patient declined further investigations for diagnosis and therapy.

According to the ROTAS study most of the improvement in visual acuity (VA) during amblyopia therapy of children aged 3 to 8 years occurs during the first 6 to 8 weeks . Sattler reported a VA gain in 11-year olds even during the second year of treatment . So far there are no standards concerning the intensity and duration of the treatment of patients older than 7 years of age. After a report on electronic monitoring of occlusion treatment in patients aged 7 to 16 years for 4 months , we now analyse whether this age group benefits from a longer-lasting treatment.In this pilot study the progression of VA was analysed in 11 patients (age range 7.18 to 15.76 years; median 11.42 years) during 12 months of occlusion therapy (types of amblyopia: 5 anisometropic, 1 strabismic, 5 combined). The daily occlusion times were recorded using the occlusion dose monitor (ODM) . At the beginning of treatment the prescription of the occlusion regime (median) was 6 h/d (range 4 to 7 h/d), the (decimal) VA 0.2 (range 0.02 to 0.63) for single and 0.16 (range 0.02 to 0.8) for crowded optotypes.The recorded occlusion time (median) was 4.4 h/d during the 12 months of treatment, the VA gain (median) was 0.4 log units for single (range 0.2 to 0.7 log units) and 0.3 for crowded optotypes (range--0.1 to 0.6). During the period of 4 to 12 months of treatment (received occlusion 4.12 h/d) the VA gain was 0.1 log units for single and for crowded optotypes. The maximum VA gain during the interval of 4 to 12 months of treatment was 0.2 log units, both single and crowded. The interocular difference for crowded VA (median) decreased from 0.9 to 0.6 log units during treatment, however only one patient achieved an interocular difference of0.2 log units.The patients presented here were able to integrate daily occlusion lasting several hours and the electronic monitoring of occlusion treatment into their daily routine over a period of 12 months. During this period the VA of all included types of amblyopia improved significantly, both from a clinical and statistical point of view. Further long-term studies using ODMs with larger groups of patients may identify factors for success of treatment, reveal the long-term stability of the improvement and contribute to a standardised treatment in this age group.

Summary In this review some aspects of endocrinological factors in cancer are discussed. The first part addresses the determination of steroid hormone receptors in cancer of the breast. The importance of steroid hormone receptor determination for the clinical management of breast cancer patients is emphasized. Current methods for receptor measurement are critically evaluated and it is concluded that rigorous standardization of tissue handling procedures and of radiochemical methodology is mandatory to improve the reliability of the presently available methods. The development of new techniques for receptor determination has been met with high expectations. Monoclonal antireceptor antibodies in principle allow the development of a radio immunoassay but are not widely available as yet. Histochemical methods for receptor localization have so far failed to meet accepted criteria of specificity. Therefore radiochemical determination at present remains the only reliable method. The second part addresses the production of peptide hormones by endocrine tumours of the gastroenteropancreatic (GEP) system and by non-neuroendocrine neoplasms. In the endocrine tumours of the GEP system a wide range of peptide hormones can be detected by immunocytochemistry, but most frequently the pattern of hormone production is comparable with that in the normal organ of origin. Clinical symptoms of excess hormone production occur much less frequently than hormone production is found by immunological methods and is usually caused by one hormone. It is proposed that these neoplasms are classified according to the hormone which causes the leading clinical symptoms and/or the hormone with the highest serum level. Peptide hormone producing cells in classical carcinoma are an intriguing phenomenon which has shed some new light on the embryological origin of neuroendocrine cells in the relevant organs and also on the histogenesis of neuroendocrine as well as non-neuroendocrine neoplasms.

Das Mykolsauregranulom in der Subcutis des Meerschweinchens ist durch eine unspezifische Fremdkorpergranulomatose mit lipophagen Reaktionen und Riesenzellen gekennzeichnet. Tuberkelstrukturen werden nicht ausgebildet. Die elektronenmikroskopischen Resultate, die farberischen und fermenthistochemischen Reaktionen (alkalische und saure Phosphatase, unspezifische Esterase, Glucose-6-Phosphat-Dehydrogenase) verweisen auf die hohe Stoffwechselaktivitat der Mykolsauregranulomatose; die Reaktionen fielen an den polynuclearen Riesenzellen durchschnittlich intensiver aus als an den einkernigen Makrophagen. Strukturell und fermenthistochemisch bestehen keine Beziehungen zur Meerschweinchentuberkulose. Autoradiographische Untersuchungen mit H3-Thymidin ergaben, das Riesenzellkerne im Rahmen der DNS-Verfugungszeit nicht oder nur ausnahmsweise markeirt sind. Nach einem Zeitintervall von 96 Std zwischen H3-Thymidin-Applikation und Granulomentnahme enthalten dagegen zahlreiche Riesenzellen markierte Zellkerne. Dieser Befund stutzt die auch elektronenmikroskopisch fundierte Annahme der Konfluenz bzw. Autophagocytose mononuclearer Elemente zur Riesenzelle. Es wird auf die Eignung des Mykolsauregranulommodells fur Fragen der formalen Genese von Riesenzellen und fur vergleichende Untersuchungen zur Dynamik des Granulationsgewebes mit moderner morphologischer Methodik hingewiesen.

Als seltener Befund bei Cushing-Syndrom wird ein Fall mit bilateraler kleinknotiger Adenomatose der Nebennierenrinde in Verbindung mit einer Involution des eigentlichen Rindengewebes beschrieben. Multizentrisch entstandene, langsam wachsende, funktionell autonome, hypersezernierende Adenome beider Nebennieren bewirken Hemmung der corticotropen Partialfunktion des Hypophysenvorderlappens und Atrophie des restlichen Rindengewebes mit der Folge einer Grosen- und Gewichtsminderung beider Nebennieren. Dieses Substrat last sich als Sonderform vom Typ der kontralateralen Rindenatrophie einerseits und von der knotigen Hyperplasie corticotropen Ursprungs andererseits abgrenzen.

Die Virulenzschadigung Isoniazid-resistenter Mycobakterien beim Meerschweinchen kommt in einem milderen Ablauf der Tuberkulose, durchschnittlich geringerem Organbefall, weitgehendem Fehlen exsudativer Veranderungen und Neigung zur Spontanheilung zum Ausdruck. Diese regressive Form der Tuberkulose unterscheidet sich von dem progressiv ablaufenden Standardtyp der Meerschweinchentuberkulose in quantitativer und qualitativer Hinsicht. Am Beispiel der Leber wird dargelegt, das sich das feingewebliche Substrat in eine spezifische epitheloidzellige und in eine unspezifische reticulumzellige Komponente, letztere in primare und sekundare Formen unterteilt. Primare Retothelknotchen werden mit Anderungen erregerbiologischer Eigenschaften in Zusammenhang gebracht, sekundare meist faserhaltige histiocytare Knotchen als rucktransformierte Epitheloidzelltuberkel gedeutet. Nicht faserbildende Epitheloid-und Reticulumzellknotchen vermogen sich reaktionslos zuruckzubilden.

It is shown that the sulphonylureas BZ 55 and D 860 are able to increase the number of mitoses of beta cells in the pancreatic islets which have previously been checked by colchicine. The effect of D 860 is 8 to 10 times larger and the effect of BZ 55 2 to 4 times larger than the normal one. These results are ascribed to the stimulated beta cell system. They are considered to be a further proof of the theory that these substances possess an increasing effect on the production and secretion of insulin. This corresponds to the fact that cortisone or glucose are also increasing the frequency of mitoses in beta cells.

The tumour syntropies of Cushing's syndrome are discussed on the basis of a case of metastatic mediastinal tumour – a small round cell undifferentiated carcinoma – followed by a Cushing syndrome. These tumoral syntropies are generally limited to the thymus, the bronchial tree and the pancreas, but may also occasionally appear elsewhere. The accompanying hyperfunction of the adrenal cortex may appear in various grades of intensity, i. e. 1) As a simple increase in the output of adrenal steroids without clinical symptoms of hyperadrenocorticism. 2) As an increase of adrenal cortex output accompanied by a hypochloraemic and hypopotassaemic alkalosis, however with no manifestation of Cushing syndrome. 3) A full blown Cushing syndrome. Either the tumour or the hyperadrenocorticism may be the primary disease. The underlying pathogenetic factor which causes the correlation between the two diseases has not yet been ascertained.

An lebensfrisch fixierten Nebennieren von Fallen mit bilateraler diffuser Rindenhyperplasie bei Cushing-Syndrom wurden planimetrische Kerngrosenbestimmungen durchgefuhrt. Sie hatten das Ergebnis einer durchschnittlichen, wenn auch nicht einheitlichen Kerngrosenzunahme in allen Zonen, vornehmlich in der Z. fasciculata und reticularis im Vergleich zur „Norm“ und zu Fallen mit sekundarer Rindenatrophie. Die Verbindung von Kernschwellung und progressiver Rindentransformation wird als symptomatisch fur eine vermehrte corticotrope Stimulierung dieser Formen von Cushing-Syndrom angesehen und entspricht im Prinzip experimentellen Befunden bei exogenem und endogenem Corticotropin-Uberangebot.

In einem Fall von M. Cushing mit bilateraler Nebennierenrindenhyperplasie wurde der immunhistologische ACTH-Nachweis in einem R-Zellenadenom des Hypophysenvorderlappens und in extraadenomatosen R-Zellen gefuhrt. Mit diesem Befund wird die Annahme der ACTH-Bildung in den R-Zellen der Adenohypophyse des Menschen am Beispiel eines klassischen Krankheitsbildes gestutzt.

A case report is given of a patient with Cushing's syndrome due to bilateral adrenocortical hyperplasia. The 17-OHCS in plasma showed only a low or none response to corticotrophin (ACTH), administered intravenously. The elevated excretion of 17-OHCS in urine exhibited a further increase following the first ACTH administration, but were not depressed significantly after 120 mg triamcinolone within four days. No reaction of urinary 17-OHCS was seen following a second ACTH-infusion. A plausible explanation for these phenomena can not be given. After bilateral adrenalectomy measurable amounts of 17-OHCS were excreted in urine, probably reflecting the steroid secretion of aberrant adrenal cortical tissue. Plasma clearance of exogenous cortisol was some-what delayed without disturbed formation of conjugated steroids. Postoperative well-being of the patient was achieved only by treatment with the 21-deoxysteroid oxylone or with triamcinolone combined with deoxycorticosterone acetate.

A case report is presented of a female patient with an invasive basophil adenoma of the anterior pituitary gland and with Cushing's syndrome. Following a review of the literature of Cushing's syndrome with expanding and invasive pituitary tumours with and without metastases we have divided these tumours into three groups: Pituitary tumours, which arise with no surgical treatment of the adrenals. Tumours of category 1, the growth of which is accelerated following adrenalectomy. Tumours whose growth is accelerated following adrenalectomy, but which are recognized clinically only after surgical treatment of the adrenals. Most of these tumours are now considered to be »hyperplasiogenic« growths, which continue to remain under the influence of hypothalamic factors. Primary autonomous, ACTH-secreting tumours are on the contrary seldom seen. Histological criteria are insufficient to differentiate between autonomous and regulated pituitary ACTH-producing tumours. It is assumed that this differentiation may be only possible by means of function-tests.

Bei Ratten wurde mit huhnervirulenten Stammen der aviaren Mycobakteriengruppe eine bilaterale verkaste Nebennierentuberkulose nach Art der isolierten Organtuberkulose erzeugt. Der tuberkulose Befall anderer Organe ist vergleichsweise minimal. Mit Ausnahme der Niere, in der exsudativ-granulierende Veranderungen vorkommen, liegt extraadrenal eine granulierende Tuberkulose vor. Nach Nebennierensubstrat, Geringfugigkeit der allgemeinen Tuberkulose und chronischem Verlauf bestehen Parallelen zum gleichen Krankheitsbild des Menschen. Zur Deutung dieses fur die Bearbeitung offener Fragen in der Pathogenese der Nebennierentuberkulose geeigneten experimentellen Modells wird ein Zusammenwirken mikro- und makroorganismischer Faktoren angenommen. Fur das Angehen der Infektion in der Nebenniere werden besondere erregerbiologische Eigenschaften gefordert. Makroorganismische Einflusse und unter diesen besonders die ortliche Corticosteronwirkung durften fur den weiteren Ablauf der Infektion und fur den Substratwandel in der Nebenniere masgeblich sein.

Mit Antiseren gegen synthetisches β1-24-Corticotropin vom Kaninchen wurden die antigenhaltigen Zellelemente im Hypophysenvorderlappen des Schweines und anderer Species (Mensch, Hund, Katze, Kaninchen) immunhistologisch dargestellt. Die Befunde sind mit jenen vergleichbar, die mit markierten Antiseren gegen hypophysares ACTH erreicht werden, aber pragnanter, da gegen Beimengungen gerichtete Antikorper entfallen. Umfarbungen mit der Perameisensaure-Alcianblau-PAS-Orange G-Technik ergaben, das die fluorescierenden Vorderlappenzellen R-Zellen entsprechen. Die Fluorescenz ist an das granulahaltige Cytoplasma gebunden und im Bereich von Golgikomplexen abgeschwacht, wahrend der Zellkern ausgespart bleibt. Fur die Immunhistologie zellgebundener corticotroper Substanzen wird die Verwendung von Antikorpern empfohlen, die durch Immunisierung mit einem chemisch definierten Antigen vom Typ der corticotrop wirksamen synthetischen Polypeptide erzeugt werden.

Bei Triturus cristatusLaurenti wird in 7 Fallen ein Wurmbefall von Epidermis und Leber beschrieben. Es handelt sich einheitlich um Nematodeneier, zum Teil mit Larven auf unterschiedlichen Entwicklungsstadien bzw. in einem Fall um einen ausgewachsenen, mannlichen, geschlechtsreifen Parasiten. Eine Spezifizierung uber die Zugehorigkeit zu einer der Untergruppen der Nematoden war im Schnittpraparat nicht sicher moglich.

Light and electron microscopic demonstration of antigens in tissue is possible by means of labelled antibodies. Direct and indirect immuno-fluorescence techniques and recently also the peroxidase method permit a broader application of this principle. The peroxidase technique has the advantage of requiring less equipment and of providing the possibility to obtain durable specimens. The fluorescence technique permits the association of fluorescence phenomena with certain tissue structures by means of secondary "staining", for instance localization of specific hormone production sites. This allows functional morphological deductions in healthy and pathological conditions. Flawless techniques and controls are required before a specific reaction can be acknowledged since auto-fluorescence phenomena as well as pseudo-specific and cross-reactions are sources of error leading to wrong conclusions. A standardization of methods must therefore be attempted.

The first case of benign angiofollicular lymphoma (Castleman) of the lower and upper eyelid is reported. The patient was a 79-year-old woman. The multifocal tumor was removed in two operations. The patient has since been free of complaints and the tumor has not recurred. This observation shows that Castleman's tumor must be included in the differential diagnosis of ophthalmologic benign and malignant lymphomas and pseudolymphomas. The diversity in nomenclature of this tumor expresses the varying pathogenetic and etiologic concepts. As in the present case, the majority of extrathoracal angiofollicular lymphomas of the hyaline-vascular type is obviously best interpreted as being hamartomas or choristomas. This does not exclude a poly-etiology, especially of the plasma cell type which is associated with general clinical symptoms.

The syndrome of pure gonadal dysgenesis (PGD) cannot always easily be distinguished from other disorders of gonadal development. Relations are evident with Turner's syndrome, females with hypoplastic ovaries, male pseudohermaphroditism, mixed gonadal dysgenesis and the vanishing testes syndrome. The case is reported of a 40 year old female with primary amenorrhea, alopecia, eunuchoid features, XY karyotype with normal breast development and sexual hair after estrogen therapy. On laparotomy streak ovaries were found at ovarian site. Pathohistological examination revealed on the left side wolffian duct remnants such as ductuli deferentes and epididymis besides sparse Leydig-(hilus-)cells and on the right side only a rudimentary fallopian tube with subendothelial accumulation of hyperplastic Leydig-(hilus-)cells. Serum-testosterone elevation above the normal female range (630 ng/dl) persisted following gonadectomy (151 ng/dl). Ectopic Leydig-(hilus-)cells were regarded responsible for the continuing testosterone production. The present case lies on borderline between PGD and mixed gonadal dysgenesis because remnants of wolffian duct derivatives suggest unilateral fetal testicular activity; classification as PGD however was justified in purely female body features and lacking evidence of testicular tissue.

The history of a 6-year-old girl with a tumor originating from thoracic spine and finally becoming resistant to surgery, radio-, and chemotherapy is reported. Tumor-biopsy material was studied by light and electron microscopy, in cell culture, by acetylcholinesterase ultracytochemistry, and by quantitative catecholamine analysis and this led to the rejection of the initial diagnosis of a neuroblastoma. Light microscopy revealed a uniform population of undifferentiated cells incompletely lobulated by broad fibrovascular septa. Using the electron microscope, cells were characterized by large intracellular pools of glycogen, little cytoplasm with an abundance of free ribosomes and a paucity of organelles. A few cells displayed desmosome-like attachment sites. Staining for specific and unspecific acetylcholinesterase was negative with light and electron microscopy, as were the results of catecholamine histofluorescence using the glyoxylic acid method. The latter result was confirmed by the negative outcome of quantitative analyses of dopamine, noradrenaline, and adrenaline with high pressure liquid chromatography nd electrochemical detection in tissue samples. Tumor cells could easily be maintained in culture for up to 4 weeks. None of a variety of treatments that are known to favor expression of neuronal characteristics in neuroblastoma cells (serum withdrawal, nerve growth factor, dbcAMP, dexamethasone) induced morphological differentiation in cultured tumor cells. On the basis of the clinical history, morphology, and of our experiments with tumor cells, the diagnosis of a so-called extraskeletal Ewing's sarcoma is most likely. Our results strengthen the view that a cell biology approach may be valuable in neuroblastoma differential diagnosis.

In the newt, Notophthalmus viridescens, hypophysectomy results in progressive atrophy of the thyroid cells to the point of irreversible degeneration. After exclusive TSH-stimulation of hypophysectomized newts, increased endocytotic activity of the follicular epithelium is observed. Prolactin stimulation under the same conditions prevents atrophy but does not result in increased cell activity, as expressed by the reduced amount of microvilli and the lack of endocytotic activity. Combined TSH- and prolactin stimulation also results in cell activation, but the activation level of exclusively TSH-stimulated cells is not reached. Although prolactin prevents cellular atrophy and degeneration of the follicular epithelium, it reduces the TSH-induced activation of the thyroid epithelium.