Classification of middle- and long-distance runners based upon their performance.

The scientific literature lacks consensus on classification of middle- and long-distance runners. This creates situations where the sample studied may not represent the target population and could produce misleading conclusions. Thus, we present an approach for a data-driven classification of middle- and long-distance runners according to their competition results. The best annual results of middle- and long-distance track runners participating at major (Olympics, World and European Championships) and national championships (Denmark, Sweden, Finland, Norway) were gathered for the 2012-2018 period. Overall, 1920 men's and 1808 women's performance results were gathered. The results were grouped accordingly. Quadratic discriminant analysis was applied to define the limits between the groups. Three basic categories could be proposed for classification: world class, international and national. Classification provides a realistic overview of performance standards and the number of athletes for different categories in middle- and long-distance track running in real-world settings. The performance-based classification provides data-driven and unified criteria for reporting standards on athletes' proficiency levels. It allows for more consistent reporting practices on the target population in research. In addition to scientific research, the classification could also be employed for a variety of practical purposes.

Tumour-cell-derived complement components C1r and C1s promote growth of cutaneous squamous cell carcinoma.

BACKGROUND: The incidence of epidermal keratinocyte-derived cutaneous squamous cell carcinoma (cSCC) is increasing worldwide. OBJECTIVES: To study the role of the complement classical pathway components C1q, C1r and C1s in the progression of cSCC. METHODS: The mRNA levels of C1Q subunits and C1R and C1S in cSCC cell lines, normal human epidermal keratinocytes, cSCC tumours in vivo and normal skin were analysed with quantitative real-time polymerase chain reaction. The production of C1r and C1s was determined with Western blotting. The expression of C1r and C1s in tissue samples in vivo was analysed with immunohistochemistry and further investigated in human cSCC xenografts by knocking down C1r and C1s. RESULTS: Significantly elevated C1R and C1S mRNA levels and production of C1r and C1s were detected in cSCC cells, compared with normal human epidermal keratinocytes. The mRNA levels of C1R and C1S were markedly elevated in cSCC tumours in vivo compared with normal skin. Abundant expression of C1r and C1s by tumour cells was detected in invasive sporadic cSCCs and recessive dystrophic epidermolysis bullosa-associated cSCCs, whereas the expression of C1r and C1s was lower in cSCC in situ, actinic keratosis and normal skin. Knockdown of C1r and C1s expression in cSCC cells inhibited activation of extracellular signal-related kinase 1/2 and Akt, promoted apoptosis of cSCC cells and significantly suppressed growth and vascularization of human cSCC xenograft tumours in vivo. CONCLUSIONS: These results provide evidence for the role of tumour-cell-derived C1r and C1s in the progression of cSCC and identify them as biomarkers and putative therapeutic targets in cSCC. What's already known about this topic? The incidences of actinic keratosis, cutaneous squamous cell carcinoma (cSCC) in situ and invasive cSCC are increasing globally. Few specific biomarkers for progression of cSCC have been identified, and no biological markers are in clinical use to predict the aggressiveness of actinic keratosis, cSCC in situ and invasive cSCC. What does this study add? Our results provide novel evidence for the role of complement classical pathway components C1r and C1s in the progression of cSCC. What is the translational message? Our results identify complement classical pathway components C1r and C1s as biomarkers and putative therapeutic targets in cSCC.

Cardiorespiratory fitness and health-related quality of life in women at risk for gestational diabetes.

This study examined the associations of cardiorespiratory fitness (CRF) and leisure-time physical activity (LTPA) with health-related quality of life (HRQoL) in women at risk for gestational diabetes mellitus (GDM). The participants were 39 women planning pregnancy with a history of GDM and/or BMI >29 kg/m2 . We assessed CRF by measuring maximal oxygen consumption (VO2max ) during incremental cycle ergometer exercise until voluntary fatigue. LTPA was self-reported, and HRQoL assessed with the SF-36 Health Survey (SF-36). The mean (SD) VO2max was 27 (6) mL·kg-1 ·min-1 , and the mean LTPA was 2.6 (1.7) h/wk. After controlling for BMI, VO2max was positively associated with the SF-36 General Health scale (ß 1.27, 95% CI: 0.09, 2.44, P=.035) and the Physical Component Summary (ß 0.48, 95% CI: 0.14, 0.82, P=.007). The General Health scale (P=.023) and the Physical Component Summary (P=.011) differed even between those with very poor and poor CRF. After controlling for BMI, LTPA was positively associated with the SF-36 Physical Functioning scale (rs =.34, P=.039), the General Health scale (ß 3.74, 95% CI: 0.64, 6.84, P=.020), and the Physical Component Summary (ß 1.13 95% CI: 0.19, 2.06, P=.020). To conclude, CRF and LTPA were positively associated with perceived general health and physical well-being in women planning pregnancy and at risk for GDM. Even a slightly better CRF would be beneficial for well-being among women with low levels of CRF.

Thermal Conductance of a Single-Electron Transistor.

We report on combined measurements of heat and charge transport through a single-electron transistor. The device acts as a heat switch actuated by the voltage applied on the gate. The Wiedemann-Franz law for the ratio of heat and charge conductances is found to be systematically violated away from the charge degeneracy points. The observed deviation agrees well with the theoretical expectation. With a large temperature drop between the source and drain, the heat current away from degeneracy deviates from the standard quadratic dependence in the two temperatures.

Association of breathing sound spectra with glottal dimensions in exercise-induced vocal cord dysfunction.

The objective of this study was to evaluate associations between the breathing sound spectra and glottal dimensions in exercise-induced vocal cord dysfunction (EIVCD) during a bicycle ergometry test. Nineteen subjects (mean age 21.8 years and range 13-39 years) with suspected EIVCD were studied. Vocal folds were continuously imaged with videolaryngoscopy and breathing sounds were recorded during the bicycle exercise test. Twelve subjects showed paradoxical movement of the vocal folds during inspiration by the end of the exercise. In seven subjects, no abnormal reactions in vocal folds were found; they served as control subjects. The glottal quotient (interarytenoid distance divided by the anteroposterior glottal distance) was calculated. From the same time period, the tracheal-vocal tract resonance peaks of the breathing sound spectra were analyzed, and stridor sounds were detected and measured. Subjects with EIVCD showed significantly higher resonance peaks during the inspiratory phase compared to the expiratory phase (p < 0.014). The glottal quotient decreased significantly in the EIVCD group (p < 0.001), but not in the control group. 8 out of 12 EIVCD patients (67%) showed stridor sounds, while none of the controls did. There was a significant inverse correlation between the frequencies of the breathing sound resonance peaks and the glottal quotient. The findings indicate that the typical EIVCD reaction of a paradoxical approximation of the vocal folds during inspiration, measured here as a decrease in the glottal quotient, is significantly associated with an increase in inspiratory resonance peaks. The findings are applicable in the documentation of EIVCD findings using videolaryngoscopy, in addition to giving clinicians tools for EIVCD recognition. However, the study is limited by the small number of subjects.

Correlated Emission Lasing in Harmonic Oscillators Coupled via a Single Three-Level Artificial Atom.

A single superconducting artificial atom can be used for coupling electromagnetic fields up to the single-photon level due to an easily achieved strong coupling regime. Bringing a pair of harmonic oscillators into resonance with the transitions of a three-level atom converts atomic spontaneous processes into correlated emission dynamics. We present the experimental demonstration of two-mode correlated emission lasing in harmonic oscillators coupled via a fully controllable three-level superconducting quantum system (artificial atom). The correlation of emissions with two different colors reveals itself as equally narrowed linewidths and quenching of their mutual phase diffusion. The mutual linewidth is more than 4 orders of magnitude narrower than the Schawlow-Townes limit. The interference between the different color lasing fields demonstrates that the two-mode fields are strongly correlated.

Average demands and most demanding passages of national-level female soccer matches: do small- and large-sided games replicate match demands?

Introduction: This study aims to (1) determine the average and most demanding passage (MDP) load of national-level female soccer matches and (2) evaluate the relationship between average and MDP load during small-sided games (SSGs), large-sided games (LSGs), and matches. Methods: A total of 37 national-level female soccer players from a single club senior team and the U18 team participated. The average and 1-, 3-, and 5-min MDP external (total, high-speed, and very-high-speed running distances, acceleration and deceleration distances, average metabolic power, and high-metabolic load distance) and internal loads (average heart rate, rate of perceived exertion) of the 29 league matches, ten 4 vs. 4 + goalkeepers SSGs, and six 8 vs. 8 + goalkeepers LSGs were analyzed by the Polar Team Pro player tracking system. Results: In matches, the external load variables during 1-, 3-, and 5-min MDPs were 167%-1,165%, 135%-504%, and 126%-359%, of match average values, respectively. In LSGs, all external load variables reached higher average values compared with those during matches, except for the very-high-speed running distance; however, no variable reached higher values of 1-min MDP compared with those during the matches. In SSGs, the average acceleration and deceleration distances were higher compared with those during the matches. Discussion: The findings from the present study suggest that LSGs and SSGs can be used to overload the average values of the selected external load variables compared with those during the matches; however, other training options must be explored to overload 1-min match MDPs.

Thermal conductance by the inverse proximity effect in a superconductor.

We study heat transport in hybrid lateral normal-metal-superconductor-normal-metal structures. We find the thermal conductance of a short superconducting wire to be strongly enhanced beyond the BCS value due to the inverse proximity effect, resulting from contributions of elastic cotunneling and crossed Andreev reflection of quasiparticles. Our measurements agree with a model based on the quasiclassical theory of inhomogeneous superconductivity in the diffusive limit.

A low-cost paper-based platform for fast and reliable screening of cellular interactions with materials.

A paper-based platform was developed and tested for studies on basic cell culture, material biocompatibility, and activity of pharmaceuticals in order to provide a reliable, robust and low-cost cell study platform. It is based upon a paper or paperboard support, with a nanostructured latex coating to provide an enhanced cell growth and sufficient barrier properties. Wetting is limited to regions of interest using a flexographically printed hydrophobic polydimethylsiloxane layer with circular non-print areas. The nanostructured coating can be substituted for another coating of interest, or the regions of interest functionalized with a material to be studied. The platform is fully up-scalable, being produced with roll-to-roll rod coating, flexographic and inkjet printing methods. Results show that the platform efficiency is comparable to multi-well plates in colorimetric assays in three separate studies: a cell culture study, a biocompatibility study, and a drug screening study. The color intensity is quantified by using a common office scanner or an imaging device and the data is analyzed by a custom computer software without the need for expensive screening or analysis equipment.

Perineurial cells coexpress genes encoding interstitial collagens and basement membrane zone components.

Perineurial cell cultures were established from the sciatic nerves of adult Wistar rats. Highly enriched cultures were studied with respect to the production of extracellular matrix components under conditions free from the influence of Schwann cells, axons, or the extracellular matrix of peripheral nerves. Indirect immunofluorescence staining revealed the presence of collagen type IV epitopes, and electron microscopy demonstrated patches of basement membrane on the perineurial cell surfaces. Collagenous fibrils with a diameter of 15-20 nm were also observed in the intracellular space. SDS-PAGE of radiolabeled medium proteins showed a pattern of bands suggesting the synthesis and secretion of fibronectin, and type I and IV collagens. Northern hybridizations revealed characteristic polymorphic mRNA transcripts corresponding to fibronectin, laminin B2 chain, as well as to the alpha-chain subunits of type I, III, and IV collagens. Furthermore, in situ hybridizations suggested expression of these genes by cultured perineurial cells without apparent heterogeneity within the cell populations. In situ hybridizations of sciatic nerve tissue from 2-wk-old rats also suggested that perineurial cells express alpha 1(I) and alpha 2(IV) collagen, as well as laminin B2 chain genes in vivo. This profile of matrix gene expression is different from that of Schwann cells, which do not synthesize fibronectin, or that of fibroblastic cells, which do not form a cell surface basement membrane. The capability of perineurial cells to express genes for the basement membrane zone and for interstitial collagens further adds to our understanding of the functional role of perineurial cells in developing and healing peripheral nerve, as well as in certain neoplastic lesions of neural origin, such as von Recklinghausen's neurofibromas.

Novel function for beta 1 integrins in keratinocyte cell-cell interactions.

We have examined the expression, localization, and function of beta 1 integrins on cultured human epidermal keratinocytes using polyclonal and monoclonal antibodies against the beta 1, alpha 2, alpha 3, and alpha 5 integrin subunits. The beta 1 polypeptide, common to all class 1 integrins, was localized primarily in areas of cell-cell contacts of cultured keratinocytes, as were alpha 2 and alpha 3 polypeptides, suggesting a possible role in cell-cell adhesion for these integrin polypeptides. In contrast, the fibronectin receptor alpha 5 subunit showed no such accumulations in regions of cell-cell contact but was more diffusely distributed in the keratinocyte plasma membrane, consistent with the absence of fibronectin at cell-cell contact sites. Colonies of cultured keratinocytes could be dissociated by treatment with monoclonal antibody specific to the beta 1 polypeptide. Such dissociation of cell-cell contacts also occurred under conditions where the monoclonal antibody had no effect on cell-substrate adhesion. Therefore, beta 1 integrin-dependent cell-cell adhesion can be inhibited without affecting other cell-adhesive interactions. Antibody treatment of keratinocytes maintained in either low (0.15 mM) or high (1.2 mM) CaCl2 also resulted in the loss of organization of intracellular F-actin filaments and beta 1 integrins, even when the anti-beta 1 monoclonal antibody had no dissociating effect on keratinocyte colonies at the higher calcium concentration. Our results indicate that beta 1 integrins play roles in the maintenance of cell-cell contacts between keratinocytes and in the organization of intracellular microfilaments. They suggest that in epithelial cells integrins can function in cell-cell interactions as well as in cell-substrate adhesion.

p38alpha and p38delta mitogen-activated protein kinase isoforms regulate invasion and growth of head and neck squamous carcinoma cells.

Recent studies indicate that the specificity of p38 mitogen-activated protein kinase (MAPK)-mediated cellular stress responses is determined by the expression pattern of the distinct p38 isoforms. Here, we have analysed the function of distinct p38 isoforms in the growth and invasion of head and neck squamous cell carcinomas (HNSCCs). Activation of p38 MAPK by arsenite resulted in inactivation of the ERK1,2 signaling pathway by dephosphorylation of MEK1,2 in primary human epidermal keratinocytes (HEKs), whereas in HNSCC cells this p38-mediated inhibition of the ERK1,2 pathway was absent. Quantitation of p38 pathway component mRNA expression in HNSCC cell lines (n=42) compared to HEKs (n=8) revealed that p38alpha and p38delta isoforms are predominantly expressed in both cell types and that MKK3 is the primary upstream activator expressed. Inhibition of endogenous p38alpha or p38delta activity by adenoviral delivery of corresponding dominant-negative p38 isoforms potently reduced MMP-13 and MMP-1 expressions, and suppressed the invasion of HNSCC cells through collagen. Dominant-negative p38alpha and p38delta inhibited squamous cell carcinoma (SCC) cell proliferation and inhibition of p38alpha activity also compromised survival of SCC cells. p38alpha and p38delta were predominantly expressed in HNSCCs (n=24) and nonneoplastic epithelium in vivo (n=6), with MKK3 being the primary upstream activator. Activation and expression of p38alpha and p38delta by tumor cells was detected in HNSCCs in vivo (n=16). Adenoviral expression of dominant-negative p38alpha or p38delta in cutaneous SCC cells potently inhibited their implantation in skin of severe combined immunodeficiency mice and growth of xenografts in vivo. Our results indicate that p38alpha and p38delta specifically promote the malignant phenotype of SCC cells by regulating cell survival, proliferation and invasion, suggesting these p38 MAPK isoforms as potential therapeutic targets in HNSCCs.

The function of the hand after operations for obstetric injuries to the brachial plexus.

Hand function was evaluated in 105 patients who had been operated on in early infancy for brachial plexus birth palsy. The mean follow-up after surgery was for 13.4 years (5.0 to 31.5). Fine sensation, stereognosis, grip and pinch strength and the Raimondi scale were recorded. Fine sensation was normal in 34 of 49 patients (69%) with C5-6 injury, 15 of 31 (48%) with C5-7 and in 8 of 25 (32%) with total injury. Loss of protective sensation or absent sensation was noted in some palmar areas of the hand in 12 of 105 patients (11%). Normal stereognosis was recorded in 88 of the 105 patients (84%), whereas only 9 of the 105 (9%) had normal grip strength. The mean Raimondi scale scores were 4.57 (3 to 5) (C5-6), 4.26 (1 to 5) (C5-7) and 2.16 (0 to 5) in patients with total injury. The location of impaired sensation was related to the distribution of the root injury. Avulsion type of injury correlated with poor recovery of hand function.

Grit blasted aggregates of hydroxyl apatite functionalized calcium carbonate in occluding dentinal tubules.

OBJECTIVES: This study aimed to investigate the effects of using hydroxyl apatite functionalized calcium carbonate (FCC) particles on occluding dentinal tubules. METHODS: Dentine specimens extracted from eighteen human molars with exposed dentinal tubules were divided into three groups (n = 6/group): a) Cut surface with smear layer; b) EDTA (smear layer removed with 17% EDTA for 1 min); and c) Grit blasted functionalized calcium carbonate (FCC) with and air pressure of 280 kPa. Microscopic dentinal tubule occlusion, tubule diameter and tubule area were evaluated using scanning electron microscopy (SEM) before and after grit blasting. Biomineralization of specimens was carried out in a simulated body fluid (SBF). Elemental analysis of occluding materials was carried out using energy-dispersive X-ray spectroscopy (EDX). X-ray diffraction (XRD) analysis was performed to demonstrate the crystal structure of the biomineralized layer on dentine. RESULTS: FCC particles showed penetration into the dentinal tubules by breakage of their original particle shape and size. EDTA treated surface had higher number and larger size tubules than those with smear layer or grit blasted (p < 0.005). SEM-EDX analysis revealed mineral precipitation of calcium phosphate on the SBF immersed dentin specimens. XRD analysis showed typical crystal structure of hydroxyl apatite for the biomineralized surface layer on dentine. CONCLUSIONS: Grit blasted FCC particles initially occluded effectively the opened dentinal tubules and biomineralization occurred in tubules primarily occluded by the FCC particles. However, in the optimal in vitro conditions in SBF, no difference between biomineralization was found between the grit blasted surface and the control surface. CLINICAL SIGNIFICANCE: Several materials and methods have been established for treatment of dentinal hypersensitivity although a golden standard treatment has not been discovered. Grit blasted functionalized calcium carbonate has a potential to occlude and remineralize exposed dentinal tubules. This could offer a more biological approach on treatment of dentin hypersensitivity.

Hybrid rf SQUID qubit based on high kinetic inductance.

We report development and microwave characterization of rf SQUID (Superconducting QUantum Interference Device) qubits, consisting of an aluminium-based Josephson junction embedded in a superconducting loop patterned from a thin film of TiN with high kinetic inductance. Here we demonstrate that the systems can offer small physical size, high anharmonicity, and small scatter of device parameters. The work constitutes a non-tunable prototype realization of an rf SQUID qubit built on the kinetic inductance of a superconducting nanowire, proposed in Phys. Rev. Lett. 104, 027002 (2010). The hybrid devices can be utilized as tools to shed further light onto the origin of film dissipation and decoherence in phase-slip nanowire qubits, patterned entirely from disordered superconducting films.

Activation of Smad signaling enhances collagenase-3 (MMP-13) expression and invasion of head and neck squamous carcinoma cells.

Squamous cell carcinoma (SCC) cells of the head and neck specifically express collagenase-3 (matrix metalloproteinase-13 (MMP-13)), the expression of which correlates with their invasion capacity. Transforming growth factor-beta (TGF-beta) enhances MMP-13 and collagenase-1 (MMP-1) expression and invasion of SCC cells via p38 mitogen-activated protein kinase. Here, we have examined the role of Smad signaling in regulating MMP-13 expression and in invasion of head and neck SCC cells. Treatment with TGF-beta resulted in activation of Smad2 and Smad3 in SCC cells, but had no effect on their proliferation or viability. Basal activation of Smad3 and p38 was noted in SCC cells without exogenous TGF-beta stimulation, and adenoviral delivery of Smad7 and dominant-negative Smad3 inhibited p38 activation in these cells. Adenoviral overexpression of Smad3 augmented the upregulatory effect of TGF-beta on MMP-13 expression by SCC cells. Disruption of Smad signaling by adenoviral expression of kinase-defective TGF-beta type I receptor (activin-receptor-like kinase-5), Smad7, and dominant-negative Smad3 potently suppressed the basal and TGF-beta-induced expression of MMP-13 and MMP-1 in SCC cells, and inhibited their basal and TGF-beta-induced invasion through Matrigel and type I collagen. Adenoviral overexpression of Smad7 in cutaneous and oral SCC cells significantly inhibited their implantation in skin of SCID mice and growth of xenografts in vivo, as compared to LacZ adenovirus-transduced control cells. Together, these results show that Smad signaling plays an important role in promoting the invasive phenotype of human head and neck SCC cells by upregulating their collagenase expression.

Effect of maternal intrahepatic cholestasis on fetal steroid metabolism.

Estriol, estriol sulfate, progesterone, and 17 neutral steroid sulfates, including estriol precursors and progesterone metabolites, were determined in 27 cord plasma samples collected after pregnancies complicated by intrahepatic cholestasis of the mother. The levels of these steroids were compared with those in the cord plasma of 42 healthy controls. In the cord plasma, the steroid profile after pregnancies complicated by maternal intrahepatic cholestasis differed greatly from that seen after uncomplicated pregnancy. Two main differences were found. In the disulfate fraction, the concentrations of two pregnanediol isomers, 5alpha-pregnane-3alpha,20alpha-diol and 5beta-pregnane-3alpha,20alpha-diol, were high after cholestasis. Other investigators have shown that, as a result of cholestasis, these pregnanediol sulfates circulate in greatly elevated amounts in the maternal plasma. Our results indicate that in cholestasis these steroids cross the placenta into the fetal compartment, where they circulate in elevated amounts as disulfates. Secondly, the concentrations of several steroid sulfates known to be synthesized by the fetus were significantly lower in the cholestasis group than in the healthy controls. This was especially true of 16alpha-hydroxydehydroepiandrosterone sulfate and 16alpha-hydroxypregnenolone sulfate. These results suggest that, in pregnancies complicated by maternal intrahepatic cholestasis, impairment of fetal steroid synthesis, and especially of 16alpha-hydroxylation, occurs in the fetal compartment.Thus, the changes in maternal steroid metabolism caused by cholestasis are reflected in the steroid profile of the fetoplacental circulation. Furthermore, maternal intrahepatic cholestasis may result in the production of some substance which crosses the placenta and affects fetal steroid metabolism.

Plasticity of integrin expression by nerve-derived connective tissue cells. Human Schwann cells, perineurial cells, and fibroblasts express markedly different patterns of beta 1 integrins during nerve development, neoplasia, and in vitro.

Strikingly selective expression patterns of beta 1, alpha 2, alpha 3, and alpha 5 integrin subunits were revealed in endoneurium, perineurium, and epineurium of fetal and adult human peripheral nerve by immunostaining with specific antibodies. The alpha 2 subunit was expressed only on Schwann cells both in fetal and adult nerve, whereas the alpha 3 epitopes were expressed exclusively in the adult tissue and were primarily present on perineurial cells. The alpha 5 epitopes were expressed only on the innermost cell layer of perineurium of fetal and adult nerve. The tumor cells within schwannomas and cutaneous neurofibromas expressed both alpha 2 and alpha 3 subunits, indicating that Schwann cells have the potential to express also the alpha 3 subunit in vivo. Cell cultures established from human fetal nerve and neurofibromas revealed expression of the alpha 2 and alpha 5 epitopes on Schwann cells, perineurial cells, and fibroblasts, whereas only Schwann cells contained the alpha 3 epitopes which were occasionally concentrated on the adjacent Schwann cells at cell-cell contacts. Our findings emphasize that nerve connective tissue cells change their profiles for expression of extracellular matrix receptors under conditions which have different regulatory control signals exerted by, for example, axons, humoral factors, or the extracellular matrix of the peripheral nerve. This plasticity may play an important role during nerve development and in neoplastic processes affecting the connective tissue compartments of peripheral nerve.

Type 1 neurofibromatosis: selective expression of extracellular matrix genes by Schwann cells, perineurial cells, and fibroblasts in mixed cultures.

Cutaneous neurofibromas, characteristic lesions of neurofibromatosis 1, are composed of an abundant extracellular matrix and nerve connective tissue-derived cell types: Schwann cells, perineurial cells, and fibroblasts. In this study, the extracellular matrix gene expression by these cells was examined under culture conditions that allowed them to be metabolically active and readily identifiable by morphologic and immunocytochemical criteria. Northern hybridizations demonstrated expression of genes for type I, III, IV, and VI collagens, as well as for fibronectin, laminin, and elastin. In situ hybridizations revealed that all three cell types expressed pro alpha 1 (I), pro alpha 2 (VI), and laminin B1 chain genes. However, fibroblasts did not contain [35S]cDNA-mRNA hybrids specific for type IV collagen, whereas both Schwann cells and perineurial cells expressed these genes. Perineurial cells and fibroblasts readily expressed the fibronectin gene whereas Schwann cells were essentially devoid of the corresponding mRNA. Perineurial cells also expressed the gene for laminin A chain. The results indicate that the extracellular matrix gene expression profiles of Schwann cells, perineurial cells, and fibroblasts are distinct: all three cell types are capable of expressing some of the genes for extracellular matrix components, such as type I and VI collagens, whereas Schwann cells and perineurial cells may have the primary role in synthesizing basement membrane zone components, type IV collagen and laminin. These observations potentially relate to the mechanisms of growth and development of human neurofibromas. The results attest to the applicability of the methodology utilized here to study other human tumors with mixed cell populations.

Collagen gene expression by cultured human skin fibroblasts. Abundant steady-state levels of type VI procollagen messenger RNAs.

Previous studies have suggested that procollagen types I and III are the major collagenous gene products of cultured human skin fibroblasts. In this study the expression of 10 different genes, encoding the subunit polypeptides for collagen types I-VI, by human skin fibroblasts in culture was analyzed by molecular hybridizations. Northern transfer analysis demonstrated the presence of specific mRNA transcripts for collagen types I, III, IV, V, and VI, but not for type II collagen. Quantitation of the abundance of these mRNAs by slot blot hybridizations revealed that type I, III, and VI procollagens were the major collagenous gene products of skin fibroblasts in culture. The mRNAs for type IV and V collagens represented only a small percentage of the total collagenous mRNA transcripts. Further analysis by in situ hybridization demonstrated that the majority of the cultured cells coexpressed the genes for type I, III, and VI procollagen pro-alpha chains. Further in situ hybridization analyses revealed the expression of type VI collagen genes in normal human skin. These data demonstrate that human skin fibroblast cultures can be used to study the transcriptional regulation of at least nine genetically distinct procollagen genes. The data further suggest that type VI collagen, in addition to types I and III, may be a major collagenous component of human skin.