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Purpose Dry eye syndrome (DES) is one of the most common diseases of the ocular surface. Affected persons suffer from different subjective complaints, with sometimes severe impairment in the quality of life. The aetiology and pathogenesis are multifactorial, multifaceted, and not yet fully understood. The present study is intended to provide deeper insights into possible triggering factors and correlating comorbidities. Materials and Methods In German ophthalmological practices, 306 persons (174 women, 132 men, age: 18 – 87 years) were interviewed by questionnaire on concomitant diseases and possible further triggering factors. DES was diagnosed by an ophthalmologist in 170 cases. The statistical comparative analysis between persons with and without DES was carried out using the chi-squared test (SPSS statistical software). Results DES occurred with significantly (p Conclusion Some of the known comorbidities and DES risk factors, e.g., computer work or depression, were confirmed. In contrast, the higher prevalence of hyperlipidaemia, hyperuricaemia, osteoporosis, nephrolithiasis, and fibroids among DES patients has not previously been reported. Additional studies should be performed on causal connections between DES and specific comorbidities.

Dry eye syndrome (DES) is one of the most common diseases of the ocular surface. Affected persons suffer from different subjective complaints, with sometimes severe impairment in the quality of life. The etiology and pathogenesis is multifactorial, multifaceted and not yet fully understood. The present work should provide deeper insights into possible triggering factors and correlating comorbidities.In German ophthalmological practices 306 persons (174 women, 132 men, age: 18-87 years) were interviewed by questionnaire regarding concomitant diseases and possible further triggering factors. DES was diagnosed by an ophthalmologist in 170 cases. The statistical comparative analysis between persons with and without DES was carried out using the Chi-square test (SPSS statistical software).DES occurred with a significantly (p0.05) increased frequency in women over 40 years of age, as well as in persons exposed to screen work, air conditioning, persons with chronic ocular inflammation, myomas (hysterectomy), dry skin, arterial hypertonicity in need of medication, cardiac arrhythmias, fatty liver, gastric ulcer, appendicitis, cholecystectomy, depression, hyperlipidemia, hyperuricemia, osteoporosis and nephrolithiasis.Some of the known comorbidities and DES-risk factors, e.g. computer work or depression were confirmed. In contrast, the higher prevalence of hyperlipidemia, hyperuricemia, osteoporosis, nephrolithiasis, and fibroids among DES-patients was described for the first time in this work. Causal connections between DES and certain comorbidities should constitute the subject of further investigation in the future. Zusammenfassung Zielsetzung: Das Trockene Auge (TA) ist eine der häufigsten Erkrankungen der Augenoberfläche. Betroffene leiden unter verschiedenen subjektiven Beschwerden, die die Lebensqualität mitunter stark beeinträchtigen. Die Ätiologie und Pathogenese sind multifaktoriell, vielschichtig und noch nicht vollständig geklärt. Die vorliegende Arbeit soll tiefere Einblicke in mögliche auslösende Faktoren und korrelierende Komorbiditäten geben.In deutschen Augenarztpraxen wurden 306 Personen (174 Frauen, 132 Männer, Alter: 18-87 Jahre) per Fragebogen zu Begleiterkrankungen und möglichen weiteren auslösenden Faktoren befragt. In 170 Fällen wurde ein TA durch einen Augenarzt diagnostiziert. Die statistische Vergleichsanalyse zwischen Personen mit und ohne TA wurde mit dem Chi-Quadrat-Test (Statistiksoftware SPSS) durchgeführt. Ergebnisse: TA traten signifikant (p0,05) häufiger bei Frauen über 40 Jahren auf sowie bei Personen, die Bildschirmarbeit und Klimaanlagen ausgesetzt waren, bei Personen mit chronischen Augenentzündungen, Myomen (Hysterektomie), trockener Haut, arterieller Hypertonie, die medikamentös behandelt werden musste, Herzrhythmusstörungen, Fettleber, Magengeschwür, Blinddarmentzündung, Cholezystektomie, Depression, Hyperlipidämie, Hyperurikämie, Osteoporose und Nephrolithiasis. Schlussfolgerung: Einige der bekannten Komorbiditäten und TA-Risikofaktoren, z. B. Bildschirmarbeit oder Depression, wurden bestätigt. Die höhere Prävalenz von Hyperlipidämie, Hyperurikämie, Osteoporose, Nephrolithiasis und Myomen bei TA-Patientinnen wurde dagegen in dieser Arbeit erstmals beschrieben. Kausale Zusammenhänge zwischen TA und bestimmten Komorbiditäten sollten in Zukunft Gegenstand weiterer Untersuchungen sein.

The identification of material parameters accurately describing the region-dependent mechanical behavior of human brain tissue is crucial for computational models used to assist, e.g., the development of safety equipment like helmets or the planning and execution of brain surgery. While the division of the human brain into different anatomical regions is well established, knowledge about regions with distinct mechanical properties remains limited. Here, we establish an inverse parameter identification scheme using a hyperelastic Ogden model and experimental data from multi-modal testing of tissue from 19 anatomical human brain regions to identify mechanically distinct regions and provide the corresponding material parameters. We assign the 19 anatomical regions to nine governing regions based on similar parameters and microstructures. Statistical analyses confirm differences between the regions and indicate that at least the corpus callosum and the corona radiata should be assigned different material parameters in computational models of the human brain. We provide a total of four parameter sets based on the two initial Poisson’s ratios of 0.45 and 0.49 as well as the pre- and unconditioned experimental responses, respectively. Our results highlight the close interrelation between the Poisson’s ratio and the remaining model parameters.The identified parameters will contribute to more precise computational models enabling spatially resolved predictions of the stress and strain states in human brains under complex mechanical loading conditions.

Purpose To date, there are many theories about tear transport through the canaliculi of the draining lacrimal system into the lacrimal sac but only few with supportive data. It is certain that the function of the lacrimal part of orbicularis oculi muscle (Horner-Duverney's muscle) is indispensable for the transport of “used” tears. However, the muscle's exact structure and the mechanisms of its functions are as yet unclear. To obtain deeper insights we undertook the present study. Methods Upper and lower canaliculi (including the entrance into the lacrimal sac) from donor cadavers were dissected. Some of the specimens were prepared for scanning electron microscopy (SEM) to analyze the course of muscle fibers surrounding the canaliculi. Others were sectioned for enzyme- (EHC) and immunohistochemistry (IHC) to learn about the distribution of slow and fast reacting muscle fibers in Horner-Duverney's muscle as well as to analyze the distribution of different neurotransmitters to learn more about the innervation of Horner-Duverney's muscle. Four tear duct systems taken from body donors were cut out en bloc after formalin fixation, serially sectioned and reconstructed using a newly developed technology for 3D reconstruction of histological serial sections named HiD® (Chimaera GmbH, Germany). Patients that had undergone dacryocystorhinostomy (DCR) were video-analyzed endonasally during active blinking, focusing on viewing the temporal wall of the lacrimal sac movement where the canaliculi penetrated the lacrimal sac. Results SEM revealed that muscle fibers of Horner-Duverney's muscle surround the vertical parts of the upper and lower canaliculus in a scissor like pattern whereas they ran in parallel to the first two thirds of the horizontal parts surrounding the respective canaliculus. Here, the muscle fibers were embedded in dense connective tissue forming a unique network. At the nasal third, muscle fibers left the canaliculi and ran to the posterior part of the fascia of the lacrimal sac and the lacrimal bone. EHC revealed that Horner-Duverney's muscle contained nearly an equal distribution of type I and type IIb muscle fibers compared to the superior rectus muscle which contains more type I and the masseter and iliopsoas muscles with more type IIb muscle fibers. IHC indicated presence of trigeminal, catecholaminergic and cholinergic nerve endings. 3D reconstructions supported the SEM data. Endonasal video analysis of patients after DCR with a nasally open lacrimal sac revealed bulging of the temporal wall of the lacrimal sac during blinking. On the basis of these findings, a modified lacrimal pump theory is proposed. Conclusion The results support the hypothesis that contraction of Horner-Duverney's muscle leads to closure of the canaliculi in their first two thirds based on the special arrangement of muscle fibers and connective tissue fibers. This causes the tear fluid in the canaliculi to be pressed/transported towards the lacrimal sac. The medial third of the vertical portions of the canaliculi, the canaliculus communis and the intrasaccal portion of the canaliculus are compressed by the shortening and thickening of the Horner-Duverney muscle from dorsal, which leads to a compression of the canaliculi lumens in this part of the system, thereby pushing the lacrimal fluid further towards the lacrimal sac. The mix of fast contracting and fatigue resistant muscle fibers is ideally suited for the blink mechanism that is complexly regulated by the nervous system.

The ocular surface is in constant interaction with the environment and with numerous pathogens. Therefore, complex mechanisms such as a stable tear film and local immune defense mechanisms are required to protect the eye. This study describes the detection, characterization, and putative role of surfactant protein G (SP-G/SFTA2) with respect to wound healing and surface activity. Bioinformatic, biochemical, and immunological methods were combined to elucidate the role of SP-G in tear film. The results show the presence of SP-G in ocular surface tissues and tear film (TF). Increased expression of SP-G was demonstrated in TF of patients with dry eye disease (DED). Addition of recombinant SP-G in combination with lipids led to an accelerated wound healing of human corneal cells as well as to a reduction of TF surface tension. Molecular modeling of TF suggest that SP-G may regulate tear film surface tension and improve its stability through specific interactions with lipids components of the tear film. In conclusion, SP-G is an ocular surface protein with putative wound healing properties that can also reduce the surface tension of the tear film.

Background In this study, we evaluate the value of novel suture material based on monofilamentous-extruded polyfluoroethylene (PTFE) compared to polypropylene (PPL) and Fiberwire (FW). Materials and methods 60 flexor tendons were harvested from fresh cadaveric upper extremities. 4–0 sutures strands were used in the PPL, FW and PTFE group. Knotting properties and mechanical characteristics of the suture materials were evaluated. A 4-strand locked cruciate (Adelaide) or a 6-strand (M-Tang) suture technique was applied as core sutures for a tendon repair. Two-way ANOVA tests were performed with the Bonferroni correction. Results Stable knotting was achieved with 5 throws with the PPL material, 7 throws for FW and 9 throws for PTFE. In the PPL group, linear tensile strength was 45.92 ± 12.53 N, in the FW group 80.11 ± 18.34 N and in the PTFE group 76.16 ± 29.10 N. FW and PTFE are significantly stronger than PPL but show no significant difference among each other. Similar results were obtained in the subgroup comparisons for different repair techniques. The Adelaide and the M-Tang knotting technique showed no significant difference. Conclusion Fiberwire showed superior handling and knotting properties in comparison to PTFE. However, PTFE allows easier approximation of the stumps. In both, M-Tang and Adelaide repairs, PTFE was equal to FW in terms of repair strength. Both PTFE and FW provide for a robust tendon repair so that early active motion regimens for rehabilitation can be applied.

ZusammenfassungIm Oktober 2020 vermeldeten die Laienpresse, aber auch einige medizinische Journale und Internetseiten, die angebliche Entdeckung einer neuen Speicheldrüse im Nasenrachen ausgehend von Positronen-Emissions-Tomografie-Computertomografie-Untersuchungen mit Liganden für das Prostata-spezifische Membranantigen (PSMA-PET/CT). Als interdisziplinäre Gruppe aus den Fachbereichen Anatomie, Pathologie, Nuklearmedizin und HNO-Heilkunde kommen wir zu der Ansicht, dass hier eine Anhäufung von kleinen Speicheldrüsen beschrieben wird. Kleine Speicheldrüsen im Nasenrachen und peritubar sind mindestens seit 1866 bekannt und immer wieder in der Literatur und in Lehrbüchern beschrieben. Das PSMA-PET/CT könnte aber geeignet sein, bei einer Bestrahlungsplanung funktionell relevante Anhäufungen von Schleimhautspeicheldrüsen, wie beispielsweise im Bereich des Nasenrachens, besser darzustellen und zu schonen. Dies sollte in klinischen Studien geprüft werden.

In October 2020, the lay press, but also some medical journals and websites reported the putative discovery of a new salivary gland in the nasopharynx based on prostate-specific membrane antigen positron emission tomography computed tomography (PSMA-PET/CT) examinations. As an interdisciplinary group from the fields of anatomy, pathology, nuclear medicine and otorhinolaryngology, we come to the view that an accumulation of minor salivary glands has been described here. Minor salivary glands in the nasopharynx and in the peritubar region have been described at least since 1866. The current description in PSMA-PET/CT does not justify the definition of a new, independent salivary gland. The PSMA-PET/CT could, however, be suitable to better protect salivary glands in the nasopharynx when planning radiation therapy. This should be evaluated in clinical trials.Im Oktober 2020 vermeldeten die Laienpresse, aber auch einige medizinische Journale und Internetseiten, die angebliche Entdeckung einer neuen Speicheldrüse im Nasenrachen ausgehend von Positronen-Emissions-Tomografie-Computertomografie-Untersuchungen mit Liganden für das Prostata-spezifische Membranantigen (PSMA-PET/CT). Als interdisziplinäre Gruppe aus den Fachbereichen Anatomie, Pathologie, Nuklearmedizin und HNO-Heilkunde kommen wir zu der Ansicht, dass hier eine Anhäufung von kleinen Speicheldrüsen beschrieben wird. Kleine Speicheldrüsen im Nasenrachen und peritubar sind mindestens seit 1866 bekannt und immer wieder in der Literatur und in Lehrbüchern beschrieben. Das PSMA-PET/CT könnte aber geeignet sein, bei einer Bestrahlungsplanung funktionell relevante Anhäufungen von Schleimhautspeicheldrüsen, wie beispielsweise im Bereich des Nasenrachens, besser darzustellen und zu schonen. Dies sollte in klinischen Studien geprüft werden.

To investigate the presence and distribution of epithelial and non-epithelial cholinergic system and cholinergic brush cells in the human lacrimal drainage system. The study was performed on fresh frozen human cadaveric samples of the lacrimal drainage system. Immunohistochemistry was performed for assessing the presence and distribution of cholinergic brush cell proteins—villin, acetylcholine synthesizing enzyme, choline acetyltransferase (ChAT); vesicular acetylcholine transporter (VAChT); components of canonical taste transduction signaling cascade, phospholipase C β2 (PLCβ2), and transient receptor potential cation channel, subfamily M, and member 5 (TRPM5). In addition, immunoreactivity to carbonic anhydrase 4 (CA4) was assessed. The immunoreactivity was scored as positive or negative and the distribution patterns in the canaliculi, lacrimal sac, and nasolacrimal duct were investigated. In addition, ultrastructural analysis was performed to ascertain the presence of brush cells by means of scanning electron microscopy (SEM). Villin revealed immunoreactivity in the superficial epithelial cells of lacrimal sac and nasolacrimal ducts. Positive immunoreactivity was also found for ChAT, VAChT, TRPM5, and PLCβ2. ChAT expression was limited to the superficial epithelial layers of the lacrimal sac epithelium. TRPM5 and PLCβ2 were expressed on the cell membranes, cytoplasm, and basolateral surfaces of the lacrimal sac epithelium and also showed strong expression in the submucosal glandular acinar cells. VAChT showed strong expression in the canaliculus and lacrimal sac and was expressed on the surface of the superficial epithelial cells and the submucosal glandular acinar cells and lining of the blood vessels. There was a uniformly negative immunoreactivity for CA4. SEM revealed single epithelial cells with dense tuft of rigid apical microvilli in the lacrimal sac and nasolacrimal ducts. This study provides a proof of principle for the presence of an intrinsic epithelial cholinergic mechanism in the lacrimal drainage system.

To quantify and compare the amounts of surfactant proteins SP-A, SP-B, SP-C and SP-D in the tear fluid collected from patients with dry eye syndrome and from individuals with a healthy ocular surface.Schirmer strips were used to collect tear fluid from both eyes of 241 volunteers (99 men, 142 women; age range: 18-87 years). Dry eye syndrome was diagnosed by ophthalmologists in 125 patients, whereas the healthy control group comprised 116 individuals. The total protein concentration was determined via Bradford assay. The relative concentration of surfactant proteins SP-A through -D was measured by enzyme-linked immuno-sorbent assay (ELISA).The mean relative concentrations of SP-A, SP-C and SP-D were significantly higher in the dry eye group as compared to the healthy controls (p0.05, one-way ANOVA). SP-B was also detected at a higher concentration in the dry eye group, but the difference to the control group was not statistically significant.The upregulation of SP-A and SP-D in the dry eye group is probably related to these proteins' known antimicrobial and immunomodulatory effects at the ocular surface. It may represent a pathophysiological response to the inflammatory condition of the ocular surface in dry eye. The upregulation of SP-B and SP-C may represent an effort of the lacrimal system to reduce surface tension and thus to counteract the increased tendency of the tear film to tear in dry eye.

Introduction Pathological formation of blood vessels plays a key role in the growth and metastasis of tumors and also in several serious ophthalmological diseases such as wet age-related macular degeneration (AMD) or diabetic retinopathy. In AMD treatment, aflibercept (tradename EYLEA®) is used to deactivate the underlying pathological neovascularisation. Aflibercept is a recombinant fusion protein which binds to vascular endothelial growth factor (VEGF) receptors, thereby inhibiting VEGF pathway activation. VEGF is one of the most important angiogenesis factors. Objective This analysis investigates lasting efficacy of aflibercept in vitro for later application as therapeutic agent against macular degeneration (AMD). Material and methods VEGF-ELISA assays were performed to investigate binding affinities at different aflibercept concentrations. The impact of VEGF on the proliferation of human umbilical vein endothelial cells (HUVEC) was investigated using proliferation assays. Moreover, time-dependent kinetic studies were performed to analyze different aflibercept storage durations with regard to its inhibitory capabilities on human VEGF. Results and conclusion Our results reveal that aflibercept significantly lowers the amount of unbound VEGF as well as the proliferation rate of HUVEC. Moreover, in contrast to specifications given by the manufacturer, aflibercept retains its full inhibitory effect up to at least 120 h after transference from the original vial into the injection syringe.

Purpose To investigate the presence and distribution patterns of 6 surfactant proteins in lacrimal drainage tissues of patients with primary acquired nasolacrimal duct (NLD) obstruction. Methods The presence and distribution of surfactant proteins (SP)-G and SP-H was first assessed in normal cadaveric lacrimal systems. The study was then performed in 10 samples of lacrimal sac and the respective NLDs obtained from patients suffering from primary acquired NLD obstruction who underwent either a dacryocystorhinostomy or a dacryocystectomy. The lacrimal sac samples were further divided into fundus and body, soon after their removal. Immunohistochemical labeling was performed for assessing the presence and distribution of SPs: SP-A, SP-B, SP-C, SP-D, SP-G/SFTA2, and SP-H/SFTA3. The results were then scored as positive or negative and the distribution pattern, if any, within the lacrimal sac and NLDs was assessed. Human lung tissues were used as controls. Results SP-H was demonstrated in the lining epithelia of the normal lacrimal drainage systems, whereas SP-G was uniformly negative. Immunohistochemical labeling revealed wide variations in the staining patterns of different SPs in different regions of the lacrimal sac and the NLD. SP-D and SP-G revealed uniformly negative immunoreactivity. Variable staining patterns were also noted between the superficial and basal layers of the lining epithelia. However, the goblet cells and intraepithelial mucous glands did not express any of the SPs. Conclusions This study provides a proof of principle for the presence of SP-H and absence of SP-G in the normal lacrimal drainage systems. In cases of primary acquired nasolacrimal duct obstruction, there were alterations or loss of SP expression in the lining epithelia of the lacrimal sac and NLDs, reflecting their possible role in the etiopathogenesis of primary acquired nasolacrimal duct obstruction.In cases of primary-acquired nasolacrimal duct obstruction, the expression of multiple surfactant proteins was either deranged or lost in the lining epithelium of the lacrimal sac and nasolacrimal ducts.

Summary Purpose Surfactant proteins (SPs) originally identified in lung tissue are important players in the innate immune system. Beyond this, they contribute to stability and rheology of gaseous or aqueous interphases. In the present study, we determined the expression and presence of SPs (A, B, C and D) in different areas of the human larynx. Methods mRNA expression of SP-A, -B, -C and -D was analyzed by means of RT-PCR in healthy samples of epiglottis, vocal and vestibular folds, subglottis and trachea. Distribution and localization of all four SPs were analyzed by Western blot and immunohistochemistry in healthy human tissue samples. Results All four SPs were detected at the mRNA- and protein level in the human larynx as well as by means of immunohistochemistry in the different tissue samples of the human larynx. Conclusion The results reveal that all four SPs are produced with different expression patterns within the human larynx. Based on the known functions, our results suggest that SPs might be involved in maintaining mucus rheology and subsequently they could be essential components for proper phonation. Moreover, the proteins seem to play a role in immune defense of the larynx.

Ophthalmologists and interventional radiologists are not the only professionals for whom diseases of the efferent tear duct system occupy centre stage; this applies also to ENT specialists involving endonasal conservative or surgical treatment. On the basis of current knowledge and taking account of results yielded by own research in recent years and of clinical aspects, we here give an overview of basic knowledge on the anatomy and physiology of the nasolacrimal system. In doing so functional aspects regarding tear transport as well as embryological and pathophysiological issues are integrated.

The practice of human and veterinary medicine is based on the science of anatomy and dissection courses are still irreplaceable in the teaching of anatomy. Embalming is required to preserve body donors, for which process formaldehyde (FA) is the most frequently used and well characterized biocidal substance. Since January 2016, a new occupational exposure limit (OEL) for FA of 0.37mg/m3 issued by the European Committee on Hazardous Substances is obligatory since FA has been classified as a human 1B carcinogen. The anatomical institutes in the German-speaking region are called upon to consolidate efforts to reduce use of FA in anatomical curricula and body donations. As a result, the Anatomische Gesellschaft (AG) has formed a "Working Group for Reduction of Formaldehyde Exposure in Dissection Courses" tasked with discussion and recommendation of measures to reduce FA. Based on the assessment of the Working Group, the AG has issued an official opinion to the effect that, at this point in time, embalming of body donors without FA completely is not feasible. Therefore, a combination of approaches are to be used to reduce FA exposure, including technical and structural (architectural) adaptations, modification of protocols for fixation and preservation as well as organizational measures. One structural measure considered unavoidable is the integration of air supply and exhaust of individual dissecting tables into the ventilation system of the anatomy building. To embalm human body donors, intra-arterial perfusion fixation with up to 4% FA and a total fluid volume of 150mL/kg body weight will suffice. For animals where body weights and biology of bodies vary widely (i.e. special needs of fixation for ruminants, large animals as horses) perfusion fixation with up to 4% FA and a quantity of fixative solution of 10-15% of the body weight may be required. Preservation of body donors in storage (immersion) can be done with 40% ethanol or in a full bath preservation containing up to 2% FA. Corpse humidification in the dissecting room is possible with 2% phenoxyethanol, in each case without FA. In veterinary anatomy, microbiological burden is often higher and therefore might lead to a need of FA in long-time storage. Compliance with the current OEL in all institutes would appear to be feasible in combination with various organizational measures.

PURPOSE To investigate the presence and distribution patterns of 6 surfactant proteins (SPs) in the human lacrimal canaliculus. METHODS The study was performed on fresh frozen cadaveric samples of canaliculi. Immunohistochemical labeling was performed for assessing the presence and distribution of SP: SP-A, SP-B, SP-C, SP-D, SP-G/SFTA2, and SP-H/SFTA3. Immunofluorescence double staining was performed using the respective fluorescein-conjugated antibodies and the results were scored as positive or negative and the distribution pattern within the canalicular system was assessed. Western blot analysis was performed on the protein content which was resolved by reducing 15% sodium dodecyl sulfate-polyacrylamide electrophoresis and bands were studied following staining with primary and secondary antibodies. Human lung tissues were used as controls. RESULTS Fluorescence double staining with 4,6-diamidino 2-pheynlindole and SPs showed strong immunostaining for SP-A, SP-B, SP-C, SP-D, and SP-H/SFTA3. The positive immunofluorescence was noticed across all the layers of the epithelium but not the subepithelial structures. The expression was noted on the surfaces and superficial cytoplasm of the superficial and deep epithelial cells. There was no expression of SP-G/SFTA2 across the canalicular system. Western blot analysis of the proteins confirmed and concurred with the immunofluorescence findings. CONCLUSIONS This study provides a proof of principle for the presence of SPs known from lungs in the canalicular system and hypothesizes their possible functions and also their potential role in the tear flow dynamics between the ocular surface and the lacrimal drainage system.

The transcription factorNKX2.1, expressed at high levels in the medial ganglionic eminence (MGE), is a master regulator of cortical interneuron progenitor development. To identify gene candidates with expression profiles similar toNKX2.1, previous transcriptome analysis of human embryonic stem cell (hESC)-derived MGE-like progenitors revealedSFTA3as the strongest candidate. Quantitative real-time PCR analysis of hESC-derived NKX2.1-positive progenitors and transcriptome data available from the Allen Institute for Brain Science revealed comparable expression patterns forNKX2.1andSFTA3during interneuron differentiationin vitroand demonstrated highSFTA3expression in the human MGE. AlthoughSFTA3has been well studied in the lung, the possible role of this surfactant protein in the MGE during embryonic development remains unexamined. To determine ifSFTA3plays a role in MGE specification,SFTA3-/- andNKX2.1 -/-hESC lines were generated using custom designed CRISPRs. We show thatNKX2.1KOs have a significantly diminished capacity to differentiate into MGE interneuron subtypes.SFTA3KOs also demonstrated a somewhat reduced ability to differentiate down the MGE-like lineage, although not as severe relative toNKX2.1deficiency. These results suggestNKX2.1andSFTA3are co-regulated genes, and that deletion ofSFTA3does not lead to a major change in the specification of MGE derivatives.

The oral cavity with the teeth and the surrounding gingival epithelium, the periodontium, the salivary glands and other structures are open to the oral environment and thus exposed to multiple microbiological and pathogenic influences. To prevent permanent inflammatory processes such as gingivitis or periodontitis an efficient defense system is essential to ensure healthy and physiological function of the oral cavity and other interacting organic systems. Surfactant proteins (SPs), originally found in pulmonary tissue are important factors of the immune system and beyond this, support the stability and rheology of gas or fluid interfaces. This study aimed to analyze the distribution of surfactant proteins by means of Western blot and immunohistochemistry in salivary glands as well as in healthy and pathological saliva. The different expression patterns of SP-A, -B, -C and -D in healthy and pathological (periodontitis) saliva were determined using ELISA quantification. One further objective of the study was the first detection of two recent discovered proteins belonging to the surfactant protein family within human salivary glands and saliva. The results of the study reveal differences in protein expression of SP-A, -B, -C and -D within healthy and pathologic saliva. The concentration of the surfactant proteins SP-A, SP-C and SP-D is increased in saliva of people suffering from periodontal diseases, whereas by contrast, SP-B shows an opposite expression pattern. Furthermore, the results evidence the presence of SP-G and SP-H within saliva and salivary glands for the first time.

The lung constantly interacts with numerous pathogens. Thus, complex local immune defence mechanisms are essential to recognise and dispose of these intruders. This work describes the detection, characterisation and three-dimensional structure of a novel protein of the lung (surfactant-associated protein 3 (SFTA3/SP-H)) with putative immunological features. Bioinformatics, biochemical and immunological methods were combined to elucidate the structure and function of SFTA3. The tissue-specific detection and characterisation was performed by using electron microscopy as well as fluorescence imaging. Three-dimensional structure generation and analysis led to the development of specific antibodies and, as a consequence, to the localisation of a novel protein in human lung under consideration of cystic fibrosis, asthma and sepsis. In vitro experiments revealed that lipopolysaccharide induces expression of SFTA3 in the human lung alveolar type II cell line A549. By contrast, the inflammatory cytokines interleukin (IL)-1β and IL-23 inhibit expression of SFTA3 in A549. Sequence- and structure-based prediction analysis indicated that the novel protein is likely to belong to the family of lung surfactant proteins. The results suggest that SFTA3 is an immunoregulatory protein of the lung with relevant protective functions during inflammation at the mucosal sites. SFTA3: a novel lung protein with putative protective and immunological functions during inflammation in lung diseases

Background: Dry eye is one of the most common eye surface disorders. Patients suffer in particular from annoying subjective symptoms that com- promise quality of life. The aim of the study was to find out when patients consult ophthalmolo- gists in Germany and what symptoms they

To analyse the effects of long-term memantine treatment on the retinal physiology and morphology of DBA/2J mice. DBA/2J (D2J) mice received i.p. injections of the NMDA receptor antagonist memantine, which protects neurons from abnormally elevated glutamate levels, twice a day over a period of 7 months. At the age of 2, 6 and 10 months, the intraocular pressure (IOP) and electroretinograms (ERGs) were measured in all treated D2J mice, in untreated D2J controls and in C57Bl/6 (B6) wild-type mice. After the last measurement at the age of 10 months, the mice were killed and the retinae and the optic nerves were analysed morphologically. The IOP increased with age in both D2J and B6 mice with a larger increase in the D2J strain. IOPs were not influenced by memantine treatment. The response amplitude of the scotopic flash ERG decreased with age in the D2J strain. This amplitude decrease, particularly that of the b-wave, was smaller in treated D2J mice. The retinae of treated D2J mice exhibited less peripheral degeneration of cone photoreceptors, and optic nerve neuropathy was less frequent. Application of the NMDA receptor antagonist memantine diminished retinal neurodegeneration in the D2J mice and had a protective effect on the b-wave amplitude of the scotopic flash ERG. This protection may occur secondarily as memantine primarily acts on retinal ganglion cells.

Backround The nasal mucosa is characterized by a multirow high prismatic ciliated epithelium representing the first barrier of the immune defense system against microbial and other environmental pathogenic influences. A number of nonspecific defense mechanisms, including the presence of lactoferrin, peroxidases, proteases, interferons, and lysozymes in nasal secretions, act to counter inflammatory processes. The surfactant proteins (SPs) known from the lungs are important components of the innate immune system. They also influence the rheology of fluids and reduce the surface tension of gas–fluid interphases. The objective of this study was to investigate the protein expression of all four SPs. A specific aim was detection and characterization of SP-C, which had previously not been confirmed in human nasal mucosa. Methods The expression of mRNA for SP-A, -B, -C and -D was investigated using reverse transcriptase polymerase chain reaction on samples of both healthy nasal mucosa and nasal mucosa altered by inflammatory processes (allergic rhinitis and chronic rhinosinusitis). The distribution of all four proteins was determined with monoclonal antibodies using Western blot analysis as well as immunohistochemical methods. Results The results show that all four SPs, including SP-C not detected before this, are nasal mucosa components. A shift was also observed in the expression behavior of the SP-A, -B, and -D in nasal mucosa with inflammatory changes. Conclusion Based on these results, SPs appear to have an important function in immunologic and rheological process of the nasal mucosa and support the prospective therapeutic use of liposomal nasal sprays.

Surfactant proteins are broadly understood to be an important structural and functional part of the lung surfactant system. These proteins influence the intra-alveolar surface tension and contribute to innate immunity of the lung. Beside the four already known surfactant proteins SP-A, SP-B, SP-C and SP-D, two novel SPs (SP-G/SFTA2 and SP-H/SFTA3) have recently been described. These show no sequential or structural similarity with the already known SPs. However, bioinformatic prediction tools suggest physicochemical properties, which have already been described for other surfactant proteins. Although it is known that SFTA2 and SFTA3 are expressed in different human tissues, their distinct functional properties remain unclear. Here, we describe the establishment of a stable expression system for recombinant SFTA3 protein synthesis in Escherichia coli. This gives rise to future experiments with the overall aim to further examine and characterize this novel protein in more detail.

Purpose: To determine whether the epithelium of the human nasolacrimal ducts contains aquaporins (AQPs), a family of membrane proteins that function as selective pores and are able to transport water, glycerol, and other small solutes across the cell plasma membrane. Methods: Expression of AQPs 0 and 1–10 in human nasolacrimal duct tissue was determined by reverse transcription polymerase chain reaction (RT-PCR). Positive PCR amplification products were verified by direct cDNA sequencing. Western blot analysis was used to detect AQPs 3–5. Antisera specific for AQPs were used in immunohistochemical analysis to determine the presence and distribution of ten AQPs (AQP 0 and 1–9) in epithelia and subepithelial glands of the nasolacrimal ducts. All samples investigated originated from human postmortem tissue. Results: In human nasolacrimal duct samples, AQPs 1, 3, 4, 5, 7, 8, 9, and 10 were identified by RTPCR. No RT-PCR products were detected for AQPs 0, 2, and 6. All AQPs, which were detected by RT-PCR, were also confirmed by direct sequencing of the cDNA. Immunohistochemical analyses revealed AQPs 1, 3, 5, 7, 8, and 9 in human nasolacrimal duct epithelium and were detected in different cells. Expression of AQP 4 could not be detected immunohistochemically but by Western blot analysis. Protein detection of AQP 10 could not be performed due to the unavailability of an appropriate antibody. Conclusions: The results suggest specific roles for AQPs in water transport through the epithelium of the nasolacrimal ducts and underline the presumption that tear fluid components are selectively absorbed in the nasolacrimal passage.

Recent investigations support the presence of human somatostatin (SS) in the excretory system of the human lacrimal gland. To get deeper insights into a possible role of SS at the ocular surface and in the lacrimal apparatus, we investigated the distribution pattern of SS and its receptors 1–5 (SSTR1-5) by means of RT-PCR, real-time RT-PCR, Western blot and immunodot blot analysis as well as immunohistochemistry in lacrimal gland, tear fluid, conjunctiva, cornea, nasolacrimal duct epithelium, and conjunctival (HCjE) and corneal (HCE) epithelial cell lines. Cell culture experiments with HCjE and HCE were performed to analyze a possible impact of SS and inflammatory mediators on the regulation of SSTR. The results confirmed the presence of SS in lacrimal gland and tear fluid, whereas it was absent at the protein level in all other tissues and cell lines investigated. Expression of SSTR1, -2, and -5 was detectable in lacrimal gland, conjunctiva, cornea, and nasolacrimal ducts. HCjE expressed only hSSTR1 and -2, and HCE revealed only SSTR2. SSTR3 and -4 were not detected in any of the analyzed samples or cell lines. In vitro on cultured immortalized HCjE cells SS leads to a concentration-dependent down-regulation of SSTR1 mRNA but does not affect SSTR2 mRNA expression. Relative expression of SSTR1 and -2 is differentially modulated by proinflammatory cytokines and bacterial components, suggesting that the expression of both receptors is immunomodulated. Our data support an autocrine and paracrine role of SS in the lacrimal system and at the ocular surface and implicate a role of SS in corneal immunology.

The hydrophobic surfactant proteins B (SP-B) and C (SP-C) are tightly bound to phospholipids. These proteins play important roles in maintaining the surface tension-lowering properties of pulmonary surfactant. Surfactant protein A (SP-A) and D (SP-D) are extremely hydrophilic and are thought to have a role in recycling surfactant and especially in improving host defense in the lung. Moreover, SP-A supports the hydrophobic surfactant proteins during surfactant sub- type assembly and inhibits secretion of lamellar bodies into the alveolar space. During recent years surfactant proteins have also been detected at locations outside the lung such as the lacrimal apparatus. In this review, the latest information regarding SP function and regulation in the human lacrimal system, the tear film and the ocular surface is summarized with regard to mucous epithelial integrity, rheological and antimicrobial properties of the tear film, tear outflow, certain disease states and possible therapeutic perspectives.

The membrane-bound enzyme 4-hydroxybenzoic acid oligoprenyltransferase (ubiA) from E. coli is crucial for the production of ubiquinone, the essential electron carrier in prokaryotic and eukaryotic organisms. On the basis of previous modeling analyses, amino acids identified as important in two putative active sites (1 and 2) were selectively mutated. All mutants but one lost their ability to form geranylated hydroxybenzoate, irrespective of their being from active site 1 or 2. This suggests either that the two active sites are interrelated or that they are in fact only one site. With the aid of the experimental results and a new structure-based classification of prenylating enzymes, a relevant 3D model could be developed by threading. The new model explains the substrate specificities and is in complete agreement with the results of site-directed mutagenesis. The high similarity of the active fold of UbiA-transferase to that of 5-epi-aristolochene synthase (Nicotiana tabacum), despite a low homology, allows a hypothesis on a convergent evolution of these enzymes to be formed.

The aim of the present study was to evaluate the regulation of membrane-anchored mucin MUC16 by proinflammatory cytokines and bacterial components at the ocular surface. Expression and distribution of MUC16 in conjunctival (HCjE) and corneal (HCE) epithelial cell lines was monitored by RT-PCR and immunohistochemistry. To determine the regulation of MUC16, cultured HCjEs and HCEs were stimulated with different cytokines, bacterial components and bacterial supernatants, and analyzed by real-time PCR, immunodot blot and immunohistochemistry. The results indicate that MUC16 is differentially regulated between HCjEs and HCEs after challenge with inflammatory mediators and suggest shedding of MUC16 from the ocular surface epithelia into the tear film. This seems to be precisely regulated. MUC16 shedding can be differentially increased and decreased, suggesting a protective function of membrane-anchored MUC16 and supporting the hypothesis that dysregulation of membrane-anchored MUC16 at the ocular surface may be involved in dry eye pathology.

The aim of the present study was to determine the possible expression of the mucin MUC16 in the lacrimal apparatus. Expression and distribution of MUC16 in lacrimal gland, accessory lacrimal glands, and nasolacrimal ducts was monitored by RT-PCR and immunohistochemistry. MUC16 was expressed and detected in all tissues investigated. Comparable to conjunctiva and cornea it was membrane-anchored in accessory lacrimal glands whereas in lacrimal gland acinar cells and columnar cells of the nasolacrimal ducts it was stored in intracytoplasmic vesicles without membrane-association. Subepithelial serous glands of the nasolacrimal ducts revealed staining of the secretion product. Intracelluar production of MUC16 is present in lacrimal gland and epithelial cells of the nasolacrimal ducts but it is not clear whether this MUC16 is secreted. MUC16 seems to be shedded or secreted from the epithelial surface of subepithelial serous glands of the nasolacrimal ducts. Our results show that MUC16 is present in the whole lacrimal apparatus. Its distribution pattern suggests different physiological functions with regard to tear film physiology and tear outflow. Moreover, the results demonstrate the existence of so far not recognized qualitative differences in the secretion product of main lacrimal gland and accessory lacrimal glands (glands of Krause).

To evaluate the expression and presence of the surfactant proteins (SP) B and C in the lacrimal apparatus at the ocular surface and in tear fluid.Expression of SP-B and SP-C was analyzed by RT-PCR in healthy lacrimal gland, conjunctiva, meibomian gland, accessory lacrimal glands, cornea, and nasolacrimal ducts. The deposition of the hydrophobic proteins SP-B and SP-C was determined by Western blot and immunohistochemistry in healthy tissues, tear fluid, and aqueous humor.The presence of both SP-B and SP-C on mRNA and protein level was evidenced in healthy human lacrimal gland, conjunctiva, cornea, and nasolacrimal ducts. Moreover, both proteins were present in tear fluid but were absent in aqueous humor. Immunohistochemical investigations revealed production of both peptides by acinar epithelial cells of the lacrimal gland and additionally by accessory lacrimal glands of the eyelid as well as epithelial cells of the conjunctiva and nasolacrimal ducts. Immunohistochemically, healthy cornea and goblet cells revealed no reactivity.Besides the recently detected surfactant-associated proteins SP-A and SP-D, our results show that SP-B and SP-C are also peptides of the tear film, the ocular surface, and the lacrimal apparatus. Based on the current knowledge of lowering surface tension in alveolar lung cells, a similar effect of SP-B and SP-C may be assumed concerning the tear film.

Purpose To evaluate the influence of somatostatin (SST) and its analog octreotid (Oct) on corneal wound healing processes. Methods The wound healing rate in C57BL/6 mice eyes under SST and Oct treatment was analyzed using an alkali-induced corneal wounding model. Effects of SST and Oct on cell proliferation, migration and quantified protein expression of vascular endothelial growth factor (VEGF) on human corneal epithelial cells (HCE, cell line) were evaluated by means of electric cell-substrate impedance sensing, scratch migration assays and ELISA. ERK1/2 and p38 phosphorylation was investigated by semi-quantitative western blot analysis. Results Ten nanograms per microliters of SST significantly accelerated the wound closure rate of corneal defects in vivo. SST and Oct had no influence on HCE cell proliferation and migration and did not activate ERK1/2 or p38 signaling in HCE cells. However, there was increased VEGF protein expression in cytosolic proteins and medium supernatants of HCE upon Oct stimulation for 24 h. One and 10 ng/ml Oct led to a 2.5-fold and 100 ng/ml Oct to a 4-fold upregulation of VEGF protein expression. Conclusion The data implicate that SST promotes corneal wound healing in a mouse model. However, using a HCE cell line in vitro , the wound healing mechanism does not seem to be supported by proliferation and migration processes or by activation of ERK1/2 and p38 signaling pathways. Other possible mechanisms could be the activation of other pathways and the induction of growth factors such as VEGF that modulate the observed corneal wound healing process.