Culturing of Melanocytes from the Equine Hair Follicle Outer Root Sheath

Hair follicles harbor a heterogeneous regenerative cell pool and represent a putative low-to-non-invasively available source of stem cells. We previously reported a technology for culturing human melanocytes from the hair follicle outer root sheath (ORS) for autologous pigmentation of tissue engineered skin equivalents. This study translated the ORS technology to horses. We de-veloped a culture of equine melanocytes from the ORS (eMORS) from equine forelock hair follicles cultured by means of an analogue human hair follicle-based in vitro methodology. The procedure was adjusted to equine physiology by addition of equine serum to the culture medium. The hair follicles were isolated by macerating forelock skin rests, enzymatically digested and subjected to air-medium-interface cultivation method. The procedure resulted in differentiated equine melanocytes, which exhibited typical morphology, presence of melanosomes, expression of cytoskeleton proteins vimentin, α-SMA, Sox2, S100ß and tyrosinase as well as tyrosinase activity followed by production of melanin. According to all assessed parameters, eMORS could be ranked as partially melanotic melanocytes. The results of the study offer an experimental base for further insight into hair follicle biology in equine and for comparative studies of hair follicles across different species

The Middle Part of the Plucked Hair Follicle Outer Root Sheath Is Identified as an Area Rich in Lineage-Specific Stem Cell Markers

Hair follicle outer root sheath (ORS) is a putative source of stem cells with therapeutic capacity. ORS contains several multipotent stem cell populations, primarily in the distal compartment of the bulge region. However, the bulge is routinely obtained using invasive isolation methods, which require human scalp tissue ex vivo. Non-invasive sampling has been standardized by means of the plucking procedure, enabling to reproducibly obtain the mid-ORS part. The mid-ORS shows potential for giving rise to multiple stem cell populations in vitro. To demonstrate the phenotypic features of distal, middle, and proximal ORS parts, gene and protein expression profiles were studied in physically separated portions. The mid-part of the ORS showed a comparable or higher NGFR, nestin/NES, CD34, CD73, CD44, CD133, CK5, PAX3, MITF, and PMEL expression on both protein and gene levels, when compared to the distal ORS part. Distinct subpopulations of cells exhibiting small and round morphology were characterized with flow cytometry as simultaneously expressing CD73/CD271, CD49f/CD105, nestin, and not CK10. Potentially, these distinct subpopulations can give rise to cultured neuroectodermal and mesenchymal stem cell populations in vitro. In conclusion, the mid part of the ORS holds the potential for yielding multiple stem cells, in particular mesenchymal stem cells

Identifying crosstalk genetic biomarkers linking a neurodegenerative disease, Parkinson’s disease, and periodontitis using integrated bioinformatics analyses

ObjectiveTo identify the genetic linkage mechanisms underlying Parkinson’s disease (PD) and periodontitis, and explore the role of immunology in the crosstalk between both these diseases.MethodsThe gene expression omnibus (GEO) datasets associated with whole blood tissue of PD patients and gingival tissue of periodontitis patients were obtained. Then, differential expression analysis was performed to identify the differentially expressed genes (DEGs) deregulated in both diseases, which were defined as crosstalk genes. Inflammatory response-related genes (IRRGs) were downloaded from the MSigDB database and used for dividing case samples of both diseases into different clusters using k-means cluster analysis. Feature selection was performed using the LASSO model. Thus, the hub crosstalk genes were identified. Next, the crosstalk IRRGs were selected and Pearson correlation coefficient analysis was applied to investigate the correlation between hub crosstalk genes and hub IRRGs. Additionally, immune infiltration analysis was performed to examine the enrichment of immune cells in both diseases. The correlation between hub crosstalk genes and highly enriched immune cells was also investigated.ResultsOverall, 37 crosstalk genes were found to be overlapping between the PD-associated DEGs and periodontitis-associated DEGs. Using clustering analysis, the most optimal clustering effects were obtained for periodontitis and PD when k = 2 and k = 3, respectively. Using the LASSO feature selection, five hub crosstalk genes, namely, FMNL1, MANSC1, PLAUR, RNASE6, and TCIRG1, were identified. In periodontitis, MANSC1 was negatively correlated and the other four hub crosstalk genes (FMNL1, PLAUR, RNASE6, and TCIRG1) were positively correlated with five hub IRRGs, namely, AQP9, C5AR1, CD14, CSF3R, and PLAUR. In PD, all five hub crosstalk genes were positively correlated with all five hub IRRGs. Additionally, RNASE6 was highly correlated with myeloid-derived suppressor cells (MDSCs) in periodontitis, and MANSC1 was highly correlated with plasmacytoid dendritic cells in PD.ConclusionFive genes (i.e., FMNL1, MANSC1, PLAUR, RNASE6, and TCIRG1) were identified as crosstalk biomarkers linking PD and periodontitis. The significant correlation between these crosstalk genes and immune cells strongly suggests the involvement of immunology in linking both diseases

Histologische Untersuchung zur Auswirkung von Diodenlaserstrahlung auf humanes Fettgewebe in vitro : eine experimentelle Studie zur Laser-assistierten Lipoplastie (LAL)

BACKGROUND: Especially plastic surgical operations using flaps for reconstruction tend to require secondary operative corrections to reduce the redundant subcutaneous tissue. Further, adipose tissue accumulations in highly difficult regions of the human body are often a problematic surgical starting point if depending on conventional liposuction techniques. This work investigates the histological effects of fatablation using a high-powered diode laser, operating in the near infrared spectrum.METHODS: For the experiments a total number of n=59 subcutaneous fat samples of the ventro-medial thigh region and the inferior abdominal region of fresh female corpses were used. Irradiation was exclusively carried out with a high-powered diode laser (“Laserline LDF-600 Diode”) operating in the near infrared spectrum of light (808/940 nm). Following the irradiation, the fat samples were histologically stained using a modified trichrome Masson-Goldner technique. Evaluation of the histological slices was performed with a special computer program (DISKUS-Mikroskopische Bilddarstellung; Version 4.25.6). All statistical analyses were performed using the SAS statistical analysis software package, version 9.1 (SAS Institute, Cary, North Carolina, USA) and the S-plus statistical analysis software system, version 6.1 (Insightful Corp., Seattle, Washington, USA). Global significance level for all statistical tests procedures conducted was chosen to alpha=5%.RESULTS: Univariate analyses were carried out using the Pearson correlation coefficient (r). Irradiance (r ranking between -0.2284 and 0.4805) respectively energy density (r ranking between -0.1325 and 0.4911) showed weak to moderate correlation to the 10 response variables. Strongest correlation was found between energy density and collateral damage as well as irradiance and collateral damage. Analysis of covariance (ANCOVA) was used for multivariate analyses with respect to the response variables. Time (p=0.0056), energy density (p=0.0333), irradiance (p=0.0440) percental depth (p<0.0001) demonstrated a statistically significant effect on the observed cavity width. In addition irradiance (p=0.0066) and energy density (p=0.0228) showed a statistically significant effect on the calculated ratio of cavity width and collateral damage (CCD-ratio). Residual analyses showed that the model assumptions were approximately valid in all 10 models. Under consideration of the whole data an irradiance range between 250 and 400 Watt/cm2 could be evaluated. This particular range demonstrated the best results regarding the ratio of cavity width and collateral damage (CCD-ratio). These findings were obtained under continuous water irrigation of the sample surface and defocused laser beam.CONCLUSION: The results of the conducted in vitro study of laser-assisted lipoplasty (LAL) show, that a selective and gentle ablation of fatty tissue using a high-powered diode laser is possible

The course of brain-derived neurotrophic factor serum concentrations in acute depressive patients during lithium augmentation of antidepressants

Einleitung Die Lithiumaugmentation (LA) ist eine der Behandlungsstrategien mit den besten Wirksamkeitsbelegen bei therapieresistenter Depression (TRD). Die BDNF-Hypothese der Depression postuliert eine kausale Beteiligung des Neurotrophins brain derived neurotrophic factor (BDNF) an der Pathophysiologie der Depression. Eine niedrigere Konzentration von BDNF im Blutserum (sBDNF) depressiver Patienten, welche unter Therapie mit Antidepressiva ansteigt, wurde in früheren Studien gezeigt. Ziel dieser Arbeit ist es erstmalig den Verlauf von sBDNF unter LA und den Zusammenhang von sBDNF mit dem klinischen Ansprechen auf die LA bei TRD in größerer Fallzahl zu untersuchen. Wir nehmen an, dass sBDNF während der LA ansteigt und ein Zusammenhang zwischen sBDNF und Therapieansprechen besteht. Da Thrombozyten einen peripheren BDNF-Speicher stellen, wird auch der Zusammenhang zwischen sBDNF und Thrombozytenkonzentration (TK) unter LA berücksichtigt. Weiterhin vergleichen wir sBDNF bei Patienten mit TRD und einer gesunden Kontrollgruppe und nehmen an, dass sich ein niedrigeres sBDNF bei Patienten mit TRD zeigt. Methode Im Rahmen eines Teilprojektes einer prospektiven, multizentrischen Kohortenstudie zur Responseprädiktion der LA bei unipolarer TRD (ICD10 F32.1-3 und F33.1-3) wurde bei 83 Patienten sBDNF und die Depressionsschwere mittels Hamilton Depression Rating Scale (HAMD-17) vor und nach vier Wochen LA bestimmt. sBDNF vor LA wurde mit dem klinischen Ansprechen als prozentuale HAMD-Wert Veränderung unter LA korreliert. In einer Subgruppe wurde eine Korrelationsanalyse zwischen TK und sBDNF durchgeführt (nTKprä = 58; nTKpost = 48; nTKprä_und_post = 47). Die sBDNF-Spiegel der TRD-Patienten wurden mit sBDNF von 80 altersgemachten gesunden Kontrollpersonen einer Vorläuferstudie verglichen. Ergebnisse sBDNF stieg unter vierwöchiger LA signifikant an (sBDNFprä: 9,21 ± 4,1 ng/ml; sBDNFpost: 9,98 ± 4,7 ng/ml; p = 0,028). Es zeigte sich eine signifikante negative Korrelation zwischen sBDNF vor LA und der prozentualen Reduktion des HAMD-Werts (p = 0,035). Die TK korrelierte vor LA mit sBDNF (p = 0,006) nicht jedoch nach LA (p>0.05). sBDNF war bei Patienten vor (p < 0,0001) und nach (p < 0,0001) LA signifikant niedriger als bei gesunden Kontrollpersonen (sBDNFKontrollen: 16,42 ± 7,40ng/ml). Schlussfolgerung Erstmalig wurde in größerer Fallzahl der Verlauf von sBDNF unter LA untersucht. Bei Patienten mit TRD zeigte sich ein signifikanter Anstieg von sBDNF unter LA. Patienten mit niedrigerem sBDNF vor LA zeigten eine stärkere Besserung der Depressionssymptome unter LA. Damit zeigt sich sBDNF als interessanter Parameter zur Responseprädiktion der LA. Diese Ergebnisse erfordern Bestätigung in zukünftigen Untersuchungen. Die vorgelegte Studie erweitert die Kenntnis zu BDNF als biologischer Parameter bei affektiven Störungen.Introduction The augmentation of Lithium (LA) is one of the therapy strategies for treatment resistant depression (TRD) with high evidence for its effectiveness. The BDNF hypothesis of depression describes a causal relationship between BDNF and the pathophysiology of depression. Low BDNF- levels in blood-serum (sBDNF) of depressed patients, which rise under application of antidepressants, are shown in former studies. The aim of this study is to investigate the course of sBDNF during LA and the relation of sBDNF with the clinical outcome in a larger number of cases. As platelets are a peripheral reservoir of sBDNF, we also considered the correlation between sBDNF and platelet count (TK). Furthermore we compare sBDNF be-tween patients with TRD with a healthy control group and assume a lower sBDNF in patients with TRD. Method As a sub-project of a prospective multicentric cohort study on response-prediction of a LA in unipolar TRD (ICD10 F32.1-3 and F33.1-3), the concentration of sBDNF and the severity of depression were measured with the Hamilton Depression Rating Scale (HAMD-17) before and after four weeks of LA in 83 patients. Correlation analyses were performed between sBDNF before LA and the treatment response in percental change in HAMD-Score. In a subgroup, correlation analyses were performed between TK and sBDNF (nTKpre=58; nTKpost=48; nTKpre_and_post=47). sBDNF of 80 age-matched healthy controls from a prior work of the same working-group was compared with sBDNF of the patients with TRD of this study. Results sBDNF increased significantly over 4 weeks with LA (sBDNFpre: 9,21 ± 4,1 ng/ml; sBDNFpost: 9,98 ± 4,7 ng/ml; p = 0,028). A significant negative correlation between sBDNF before LA and the percental change of the HAMD-score was found (p = 0,035). We found a correlation between sBDNF and TK before LA (p = 0,006), but not after LA (p > 0,05). Patients with TRD had significantly lower sBDNF before (p < 0,0001) and after (p < 0,0001) LA than the healthy control group (sBDNFcontrols: 16,42 ± 7,40ng/ml; 16,42 ± 7,40ng/ml). Conclusion For the first time the course of sBDNF was examined in a larger number of patients receiving a LA. Patients with TRD showed an increase of sBDNF during LA. Patients with lower sBDNF before LA showed a better improvement of depressive symptoms during LA. Therefore, sBDNF is an interesting parameter for response prediction of a LA. These results need confirmation in future studies. This work extends the knowledge regarding sBDNF as a bio-logical parameter in affective disorders

Histologische Untersuchung zur Auswirkung von Diodenlaserstrahlung auf humanes Fettgewebe in vitro : eine experimentelle Studie zur Laser-assistierten Lipoplastie (LAL)

BACKGROUND: Especially plastic surgical operations using flaps for reconstruction tend to require secondary operative corrections to reduce the redundant subcutaneous tissue. Further, adipose tissue accumulations in highly difficult regions of the human body are often a problematic surgical starting point if depending on conventional liposuction techniques. This work investigates the histological effects of fatablation using a high-powered diode laser, operating in the near infrared spectrum.METHODS: For the experiments a total number of n=59 subcutaneous fat samples of the ventro-medial thigh region and the inferior abdominal region of fresh female corpses were used. Irradiation was exclusively carried out with a high-powered diode laser (“Laserline LDF-600 Diode”) operating in the near infrared spectrum of light (808/940 nm). Following the irradiation, the fat samples were histologically stained using a modified trichrome Masson-Goldner technique. Evaluation of the histological slices was performed with a special computer program (DISKUS-Mikroskopische Bilddarstellung; Version 4.25.6). All statistical analyses were performed using the SAS statistical analysis software package, version 9.1 (SAS Institute, Cary, North Carolina, USA) and the S-plus statistical analysis software system, version 6.1 (Insightful Corp., Seattle, Washington, USA). Global significance level for all statistical tests procedures conducted was chosen to alpha=5%.RESULTS: Univariate analyses were carried out using the Pearson correlation coefficient (r). Irradiance (r ranking between -0.2284 and 0.4805) respectively energy density (r ranking between -0.1325 and 0.4911) showed weak to moderate correlation to the 10 response variables. Strongest correlation was found between energy density and collateral damage as well as irradiance and collateral damage. Analysis of covariance (ANCOVA) was used for multivariate analyses with respect to the response variables. Time (p=0.0056), energy density (p=0.0333), irradiance (p=0.0440) percental depth (p<0.0001) demonstrated a statistically significant effect on the observed cavity width. In addition irradiance (p=0.0066) and energy density (p=0.0228) showed a statistically significant effect on the calculated ratio of cavity width and collateral damage (CCD-ratio). Residual analyses showed that the model assumptions were approximately valid in all 10 models. Under consideration of the whole data an irradiance range between 250 and 400 Watt/cm2 could be evaluated. This particular range demonstrated the best results regarding the ratio of cavity width and collateral damage (CCD-ratio). These findings were obtained under continuous water irrigation of the sample surface and defocused laser beam.CONCLUSION: The results of the conducted in vitro study of laser-assisted lipoplasty (LAL) show, that a selective and gentle ablation of fatty tissue using a high-powered diode laser is possible

Chronic stridor in a toddler after ingestion of a discharged button battery: a case report

Oftring ZS, Mehrtens DM, Mollin J, Hamelmann E, Gaus S. Chronic stridor in a toddler after ingestion of a discharged button battery: a case report. BMC Pediatrics. 2024;24(1): 246.Background Button battery (BB) ingestions (BBI) are increasingly prevalent in children and constitute a significant, potentially life-threatening health hazard, and thus a pediatric emergency. Ingested BBs are usually charged and can cause severe symptom within 2 h. Discharged BBs ingestion is very rare and protracted symptom trajectories complicate diagnosis. Timely imaging is all the more important. Discharged BBs pose specific hazards, such as impaction, and necessitate additional interventions.Case presentation We present the case of a previously healthy 19-month-old girl who was admitted to our pediatric university clinic in Germany for assessment of a three-month history of intermittent, mainly inspiratory stridor, snoring and feeding problems (swallowing, crying at the sight of food). The child's physical examination and vital signs were normal. Common infectious causes, such as bronchitis, were ruled out by normal lab results including normal infection parameters, negative serology for common respiratory viruses, and normal blood gas analysis, the absence of fever or pathological auscultation findings. The patient's history contained no evidence of an ingestion or aspiration event, no other red flags (e.g., traveling, contact to TBC). Considering this and with bronchoscopy being the gold standard for foreign body (FB) detection, an x-ray was initially deferred. A diagnostic bronchoscopy, performed to check for airway pathologies, revealed normal mucosal and anatomic findings, but a non-pulsatile bulge in the trachea. Subsequent esophagoscopy showed an undefined FB, lodged in the upper third of the otherwise intact esophagus. The FB was identified as a BB by a chest X-ray. Retrieval of the battery proved extremely difficult due to its wedged position and prolonged ingestion and required a two-stage procedure with consultation of Ear Nose Throat colleagues. Recurring stenosis and regurgitation required one-time esophageal bougienage during follow-up examinations. Since then, the child has been asymptomatic in the biannual endoscopic controls and is thriving satisfactorily.Conclusion This case describes the rare and unusual case of a long-term ingested, discharged BB. It underscores the need for heightened vigilance among healthcare providers regarding the potential hazards posed by discharged BBIs in otherwise healthy children with newly, unexplained stridor and feeding problems. This case emphasizes the critical role of early diagnostic imaging and interdisciplinary interventions in ensuring timely management and preventing long-term complications associated even to discharged BBs

Decision-Making in the Pediatric Emergency Department&mdash;A Survey of Guidance Strategies among Residents

(1) Introduction: Working in an emergency department requires fast and straightforward decisions. Therefore, decision guidance represents an essential tool for successful patient-centered care. Beyond the residents&rsquo; own knowledge and experience, printed books have been the primary source of information in the past. The aim of this study was to discover which strategies current residents use the most and to identify alternative quick reference strategies in the digital age. (2) Materials and Methods: This study analyzed the responses of a short questionnaire directed at 41 residents in a single pediatric emergency department (32 pediatric and 9 pediatric surgery residents) over a period of one month. (3) Results: Thirty-three (80.5%) residents answered the entire questionnaire. Strikingly, responses indicated that printed books are still pivotal in guiding decision-making. In addition, the acquisition of information via computers or smartphones plays an increasing role. However, the opinion and council of the attending physician is still of great value to the residents and is not to be underestimated. Overall, most of the residents would prefer to have access to a specially designed smartphone application. (4) Conclusions: Certainty and validity are essential in decision-making in a pediatric emergency department. Although printed books and attending physicians are still considered as reliable sources of information, internet-based information plays an increasing role. In order to provide the best up-to-date and most recent information, a validated and consistently updated smartphone application could be a useful option

Exploring Monological and Dialogical Phases in Naturally Occurring Explanations

Fisher JB, Lohmer V, Kern F, Barthlen W, Gaus S, Rohlfing KJ. Exploring Monological and Dialogical Phases in Naturally Occurring Explanations. KI - Künstliche Intelligenz. 2022.Recent approaches to Explainable AI (XAI) promise to satisfy diverse user expectations by allowing them to steer the interaction in order to elicit content relevant to them. However, little is known about how and to what extent the explainee takes part actively in the process of explaining. To tackle this empirical gap, we exploratively examined naturally occurring everyday explanations in doctor–patient interactions (N = 11). Following the social design of XAI, we view explanations as emerging in interactions: first, we identified the verbal behavior of both the explainer and the explainee in the sequential context, which we could assign to phases that were either monological or dialogical; second, we investigated in particular who was responsible for the initiation of the different phases. Finally, we took a closer look at the global conversational structure of explanations by applying a context-sensitive model of organizational jobs, thus adding a third layer of analysis. Results show that in our small sample of conversational explanations, both monological and dialogical phases varied in their length, timing of occurrence (at the early or later stages of the interaction) and their initiation (by the explainer or the explainee). They alternated several times in the course of the interaction. However, we also found some patterns suggesting that all interactions started with a monological phase initiated by the explainer. Both conversational partners contributed to the core organizational job that constitutes an explanation. We interpret the results as an indication for naturally occurring everyday explanations in doctor–patient interactions to be co-constructed on three levels of linguistic description: (1) by switching back and forth between monological to dialogical phases that (2) can be initiated by both partners and (3) by the mutual accomplishment and thus responsibility for an explanation’s core job that is crucial for the success of the explanation. Because of the explorative nature of our study, these results need to be investigated (a) with a larger sample and (b) in other contexts. However, our results suggest that future designs of artificial explainable systems should design the explanatory dialogue in such a way that it includes monological and dialogical phases that can be initiated not only by the explainer but also by the explainee, as both contribute to the core job of explicating procedural, clausal, or conceptual relations in explanations

Culturing of Melanocytes from the Equine Hair Follicle Outer Root Sheath

Hair follicles harbor a heterogeneous regenerative cell pool and represent a putative low-to-non-invasively available source of stem cells. We previously reported a technology for culturing human melanocytes from the hair follicle outer root sheath (ORS) for autologous pigmentation of tissue engineered skin equivalents. This study translated the ORS technology to horses. We de-veloped a culture of equine melanocytes from the ORS (eMORS) from equine forelock hair follicles cultured by means of an analogue human hair follicle-based in vitro methodology. The procedure was adjusted to equine physiology by addition of equine serum to the culture medium. The hair follicles were isolated by macerating forelock skin rests, enzymatically digested and subjected to air-medium-interface cultivation method. The procedure resulted in differentiated equine melanocytes, which exhibited typical morphology, presence of melanosomes, expression of cytoskeleton proteins vimentin, α-SMA, Sox2, S100ß and tyrosinase as well as tyrosinase activity followed by production of melanin. According to all assessed parameters, eMORS could be ranked as partially melanotic melanocytes. The results of the study offer an experimental base for further insight into hair follicle biology in equine and for comparative studies of hair follicles across different species