Effect of two shampoo formulations on the prokaryotic and eukaryotic microbiota composition of the human scalp.

OBJECTIVE: The human scalp is characterized by a moderately diverse microbial community, comprising prokaryotic (bacteria) and eukaryotic (fungi) members. Although the details are far from being fully understood, the human scalp microbiota is implicated in several scalp disorders, in particular dandruff formation. Hence, the protection of an intact and diverse scalp microbiota can be regarded as a quality criterion for hair and scalp care formulations. In this study, we investigated the influence of two commercially available, non-antimicrobial shampoo formulations on the structure of the scalp microbiota. METHODS: Scalp microbiota samples, obtained by swab sampling from two cohorts of probands (n = 25, each), were analysed before and after daily use of two different shampoo formulations for 2 weeks, respectively. A polyphasic approach was used, comprising quantitative cultivation of bacteria and fungi on selective media as well as sequencing of PCR-amplified 16S rRNA and 18S rRNA genes, respectively. RESULTS: All analyses revealed a microbiota composition typical for the human scalp. While in particular fungal germ numbers increased significantly during the treatments, overall bacterial and fungal community composition was not affected, based on alpha- and beta-diversity measures. However, we observed an increase in structural bacterial diversity with the age of the probands. CONCLUSIONS: Over an application period of 2 weeks, the investigated shampoo induced quantitative but no qualitative changes in the scalp microbial community structure of the investigated probands, suggesting no adverse but rather preserving or even stimulating effects of the underlying formulations on the scalp microbiota. Further investigation will have to clarify if this is also true for longer application periods and if the formulations might affect community functionality, for example microbial gene expression, rather than community composition.

OBJECTIF: Le cuir chevelu humain se caractérise par une communauté microbienne modérément diversifiée, comprenant des membres procaryotes (bactéries) et eucaryotes (champignons). Bien que l'on soit loin de comprendre totalement les détails, le microbiote du cuir chevelu humain est impliqué dans différents troubles du cuir chevelu, en particulier la formation de pellicules. La protection du microbiote du cuir chevelu intact et diversifié peut être considérée comme un critère de qualité pour les formulations de soins pour les cheveux et le cuir chevelu. Dans cette étude, nous avons examiné l'influence de deux formulations de shampooing non antimicrobien disponibles dans le commerce sur la structure du microbiote du cuir chevelu. MÉTHODES: Des échantillons de microbiote du cuir chevelu, obtenus par écouvillonnage dans deux cohortes de proposants (n = 25 dans chaque cohorte), ont été analysés respectivement avant et après l'utilisation quotidienne de deux formulations de shampooing pendant deux semaines. Une approche en plusieurs phases a été utilisée, dont une culture quantitative de bactéries et de champignons sur des milieux sélectifs et un séquençage respectivement des gènes de l'ARN ribosomique 16S et de l'ARN ribosomique 18S amplifiés par PCR. RÉSULTATS: Toutes les analyses ont révélé une composition du microbiote typique pour le cuir chevelu humain. Bien que le nombre de germes fongiques en particulier ait augmenté significativement pendant les traitements, la composition globale des communautés bactériennes et fongiques n'a pas été affectée, d'après les mesures de diversité alpha et bêta. Cependant, nous avons observé une augmentation de la diversité bactérienne structurelle avec l'âge des proposants. CONCLUSIONS: Sur une période d'utilisation de deux semaines, le shampooing étudié a induit des modifications quantitatives, mais pas qualitatives, de la structure des communautés microbiennes du cuir chevelu des proposants étudiés, ce qui suggère qu'il n'y a pas d'effets indésirables, mais qu'il y a des effets de préservation, voire de stimulation, des formulations sous-jacentes sur le microbiote du cuir chevelu. Des recherches supplémentaires devront clarifier si cela s'avère également pour des périodes d'utilisation plus longues et si les formulations peuvent affecter la fonctionnalité des communautés, par exemple, l'expression des gènes microbiens, plutôt que la composition des communautés.

Measuring lung mechanics of expiratory tidal breathing with non-invasive breath occlusion.

BACKGROUND AND OBJECTIVE: Lung mechanics measurements provide clinically useful information about disease progression and lung health. Currently, there are no commonly practiced methods to non-invasively measure both resistive and elastic lung mechanics during tidal breathing, preventing the important information provided by lung mechanics from being utilised. This study presents a novel method to easily assess lung mechanics of spontaneously breathing subjects using a dynamic elastance, single-compartment lung model. METHODS: A spirometer with a built-in shutter was used to occlude expiration during tidal breathing, creating exponentially decaying flow when the shutter re-opened. The lung mechanics measured were respiratory system elastance and resistance, separated from the exponentially decaying flow, and interrupter resistance calculated at shutter closure. Progressively increasing resistance was added to the spirometer mouthpiece to simulate upper airway obstruction. The lung mechanics of 17 healthy subjects were successfully measured through spirometry. RESULTS: N = 17 (8 female, 9 male) healthy subjects were recruited. Measured decay rates ranged from 5 to 42/s, subjects with large variation of decay rates showed higher muscular breathing effort. Lung elastance measurements ranged from 3.9 to 21.2 cmH[Formula: see text]O/L, with no clear trend between change in elastance and added resistance. Resistance calculated from decay rate and elastance ranged from 0.15 to 1.95 cmH[Formula: see text]Os/L. These very small resistance values are due to the airflow measured originating from low-resistance areas in the centre of airways. Occlusion resistance measurements were as expected for healthy subjects, and increased as expected as resistance was added. CONCLUSIONS: This test was able to identify reasonable dynamic lung elastance and occlusion resistance values, providing new insight into expiratory breathing effort. Clinically, this lung function test could impact current practice. It does not require high levels of cooperation from the subject, allowing a wider cohort of patients to be assessed more easily. Additionally, this test can be simply implemented in a small standalone device, or with standard lung function testing equipment.

Eye-Catching Microbes-Polyphasic Analysis of the Microbiota on Microscope Oculars Verifies Their Role as Fomites.

Microscopes are used in virtually every biological and medical laboratory. Previous cultivation-based studies have suggested that direct contact with microscope eyepieces increases the risk of eye infections. To obtain a deeper insight into the microbiota on oculars, we analysed 10 recently used university microscopes. Their left oculars were used for a cultivation-based approach, while the right oculars served for massive gene sequencing. After cleaning with isopropyl alcohol, the oculars were re-sampled and analysed again. All oculars were found to be contaminated with bacteria, with a maximum load of 1.7 × 103 CFU cm-2. MALDI Biotyping revealed mainly Cutibacterium (68%), Staphylococcus (14%) and Brevibacterium (10%), with the most abundant species being Cutibacterium acnes (13%) and Staphylococcus capitis (6%). Cleaning reduced the microbial load by up to 2 log scales. Within 10 uncleaned and 5 cleaned samples, 1480 ASVs were assigned to 10 phyla and 262 genera. The dominant genera before cleaning were Cutibacterium (78%), Paracoccus (13%), Pseudomonas (2%) and Acinetobacter (1%). The bacteriota composition on the cleaned oculars was similar; however, it probably largely represented dead bacteria. In summary, used oculars were significantly contaminated with skin and environmental bacteria, including potential pathogens. Regular cleaning is highly recommended to prevent eye and skin infections.

Site-specific molecular analysis of the bacteriota on worn spectacles.

Regularly touched surfaces are usually contaminated with microorganisms and might be considered as fomites. The same applies for spectacles, but only little is known about their microbial colonization. Previous cultivation-based analyses from our group revealed a bacterial load strongly dominated by staphylococci. To better account for aerotolerant anaerobes, slow growing and yet-uncultivated bacteria, we performed an optimized 16S rRNA gene sequencing approach targeting the V1-V3 region. 30 spectacles were swab-sampled at three sites, each (nosepads, glasses and earclips). We detected 5232 OTUs affiliated with 19 bacterial phyla and 665 genera. Actinobacteria (64%), Proteobacteria (22%), Firmicutes (7%) and Bacteroidetes (5%) were relatively most abundant. At genus level, 13 genera accounted for 84% of the total sequences of all spectacles, having a prevalence of more than 1% relative abundance. Propionibacterium (57%), Corynebacterium (5%), Staphylococcus (4%), Pseudomonas, Sphingomonas and Lawsonella (3%, each) were the dominant genera. Interestingly, bacterial diversity on the glasses was significantly higher compared to nosepads and earclips. Our study represents the first cultivation-independent study of the bacteriota of worn spectacles. Dominated by bacteria of mostly human skin and epithelia origin and clearly including potential pathogens, spectacles may play a role as fomites, especially in clinical environments.

Regulation of WNT5A and WNT11 during MSC in vitro chondrogenesis: WNT inhibition lowers BMP and hedgehog activity, and reduces hypertrophy.

Re-directing mesenchymal stromal cell (MSC) chondrogenesis towards a non-hypertrophic articular chondrocyte-(AC)-like phenotype is important for improving articular cartilage neogenesis to enhance clinical cartilage repair strategies. This study is the first to demonstrate that high levels of non-canonical WNT5A followed by WNT11 and LEF1 discriminated MSC chondrogenesis from AC re-differentiation. Moreover, ß-catenin seemed incompletely silenced in differentiating MSCs, which altogether suggested a role for WNT signaling in hypertrophic MSC differentiation. WNT inhibition with the small molecule IWP-2 supported MSC chondrogenesis according to elevated proteoglycan deposition and reduced the characteristic upregulation of BMP4, BMP7 and their target ID1, as well as IHH and its target GLI1 observed during endochondral differentiation. Along with the pro-hypertrophic transcription factor MEF2C, multiple hypertrophic downstream targets including IBSP and alkaline phosphatase activity were reduced by IWP-2, demonstrating that WNT activity drives BMP and hedgehog upregulation, and MSC hypertrophy. WNT inhibition almost matched the strong anti-hypertrophic capacity of pulsed parathyroid hormone-related protein application, and both outperformed suppression of BMP signaling with dorsomorphin, which also reduced cartilage matrix deposition. Yet, hypertrophic marker expression under IWP-2 remained above AC level, and in vivo mineralization and ectopic bone formation were reduced but not eliminated. Overall, the strong anti-hypertrophic effects of IWP-2 involved inhibition but not silencing of pro-hypertrophic BMP and IHH pathways, and more advanced silencing of WNT activity as well as combined application of IHH or BMP antagonists should next be considered to install articular cartilage neogenesis from human MSCs.