Revista complutense de educación

Resumen basado en el de la publicaciónSe lleva a cabo una revisión general del procedimiento cloze, procedimiento que es ampliamente conocido y utilizado como instrumento de evaluación de la lectura en los países de habla inglesa pero que apenas es conocido y empleado en España. Dicha revisión hace referencia tanto a los aspectos metodológicos relacionados con dicho procedimiento como a los distintos usos para los que puede emplearse en el campo de la evaluación de la lectura.ES

An Allelic/Nonallelic Series of BMP-Deficient Limbs

<div><p>(A–D) <i>Prx1::cre</i> efficiently recombines <i>Bmp2</i> and <i>Bmp4</i> conditional alleles in the limbs. <i>Bmp2</i> (A and B) and <i>Bmp4</i> (C and D) whole mount mRNA in situ hybridization in the limb. Wild-type (A) and <i>Bmp2^C/C</i>; <i>Prx1::cre</i> (B) are forelimbs from E10.5 mouse embryos. Mesenchymal expression [asterisk (A)] of <i>Bmp2</i> is abolished in <i>Bmp2^C/C</i>; <i>Prx1::cre</i> embryo while the AER expression [black arrow (A and B)] of <i>Bmp2</i> persists. Note that pink staining in the central region of the limb bud in (B) is nonspecific background. Wild-type (C) and <i>Bmp4^C/C</i>; <i>Prx1::cre</i> (D) are forelimbs from E10.5 mouse embryos. Mesenchymal expression [red arrow (C)] of <i>Bmp4</i> is abolished in <i>Bmp4^C/C</i>; <i>Prx1::cre</i> embryo while the AER expression [black arrow (C and D)] of <i>Bmp4</i> persists.</p><p>(E–T) Depletion of BMP2 and BMP4 together causes severe limb skeletal defects. (E–T) Whole mount skeletons from newborn animals stained with Alcian blue and Alizarin red. (E–L) Forelimbs, (M–T) hindlimbs. (E and M) Wild-type, (F and N) <i>Bmp2^C/C</i>; <i>Prx1::cre</i>, (G and O) <i>Bmp4^C/C</i>; <i>Prx1::cre</i>, (H and P) <i>Bmp7</i> ^{−<i>/</i>−}, (I and Q) <i>Bmp2^C/C</i>; <i>Bmp4^+/C</i>; <i>Prx1::cre</i>, (J and R) <i>Bmp2^+/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i>, (K and S) <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre,</i> (L and T) <i>Bmp2^C/C</i>, <i>Bmp7</i> ^{−<i>/</i>−}; <i>Prx1::cre.</i> Thin red arrow in (F), (I), and (L), defective scapula; thick red arrow in (T), failure of fibula to articulate with knee, and thick black arrow in (L) and (T), missing phalanx in digit III.</p></div

TNFα promotes proliferation of human synovial MSCs while maintaining chondrogenic potential

<div><p>Synovial mesenchymal stem cells (MSCs) are a candidate cell source for cartilage and meniscus regeneration. If we can proliferate synovial MSCs more effectively, we can expand clinical applications to patients with large cartilage and meniscus lesions. TNFα is a pleiotropic cytokine that can affect the growth and differentiation of cells in the body. The purpose of this study was to examine the effect of TNFα on proliferation, chondrogenesis, and other properties of human synovial MSCs. Passage 1 human synovial MSCs from 2 donors were cultured with 2.5 x 10⁻¹²~10⁻⁷ g/ml, 10 fold dilution series of TNFα for 14 days, then the cell number and colony number was counted. The effect of the optimum dose of TNFα on proliferation was also examined in synovial MSCs from 6 donors. Chondrogenic potential of synovial MSCs pretreated with TNFα was evaluated in 6 donors. The expressions of 12 surface antigens were also examined in 3 donors.2.5 ng/ml and higher concentration of TNFα significantly increased cell number/dish and cell number/colony in both donors. The effect of 25 ng/ml TNFα was confirmed in all 6 donors. There was no significant difference in the weight, or amount of glycosaminoglycan and DNA of the cartilage pellets between the MSCs untreated and MSCs pretreated with 25 ng/ml TNFα. TNFα decreased expression rate of CD 105 and 140b in all 3 donors. TNFα promoted proliferation of synovial MSCs with increase of cell number/ colony. Pretreatment with TNFα did not affect chondrogenesis of synovial MSCs. However, TNFα affected some properties of synovial MSCs.</p></div

Chondrogenesis Starts and Proceeds Normally Even in the Absence of BMP2 and BMP4

<div><p>(A and B) Whole mount skeletons from E13.5 embryos that are stained with alcian blue. (A) Wild-type embryo, (B) <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> embryo. Black arrow in (B) shows the fusion of the zeugopod and stylopod.</p><p>(C and D) Hematoxylin and eosin–stained sagittal sections of humeri from E13.5 wild-type (C) and <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> (D) embryos. Thick red arrow in (C) shows the hypertrophic region.</p><p>(E and F) Sagittal sections of wild-type (E) and <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> (F) humeri from E15.5 embryos were hybridized with digoxigenin-labeled antisense rioprobes for <i>ColX</i>.</p><p>(G and H) Sagittal sections of forelimbs from E13.5 wild-type and BMP2, BMP4–deficient embryos, respectively, stained with radioactive riboprobes for <i>Bmp7</i> mRNA. (G′) and (H′) show the bright field views of (G) and (H), respectively.</p></div

Comparison of chondrogenic potential of synovial MSCs pretreated with or without TNFα.

<p>(A) Experimental design. Synovial MSCs pretreated with 25 ng/ml TNFα or without TNFα (Control) were harvested, pelleted, and cultured in the chondrogenic medium without TNFα for 21 days. (B) Macroscopic and histological features of cartilage pellets. For histology, the sections were stained with safranin-o (C) Diameter and weight of the cartilage pellets. Average values are shown (n = 6, *p<0.05 by Wilcoxon signed-ranks test).</p

Defects in Bone Formation in the Absence of BMP2 and BMP4

<div><p>Toluidine blue staining of sagittal sections of distal femurs.</p><p>(A–E) Control femurs at 1 wk (A) and 3 wks (D) of age. Boxed areas in (A) are enlarged in (B) and (C). Boxed area in (D) is enlarged in (E). Blue arrows in (C) point to osteoblast cells lining the surface of cortical bone. Pink arrows in (G) show similar cells to those in (C) in <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> femurs. Red star in (A) and (D) marks the secondary ossification center. Pale blue–stained tissue in (E), marked by red arrows, is trabecular bone. bm, marks the bone marrow cavity in (D).</p><p>(F–J) <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> femurs at 1 wk (F) and 3 wks (J) of age. Boxed areas in (F) are enlarged in (G), (H), and (I). (A), (D), (F), and (J) are shown with the same magnification. Note that there are defects in bone formation but no defect in osteoclast mediated bone resorption. Mineralized tissue in the mid shaft of femur is almost resorbed at 3 wks. (H) Fibroblast-like cells present in the bone shaft of <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> mouse. Red star in (I) shows the osteoclasts invading the <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> femurs at 1 wk of age. s and m mark soft tissues and muscle in (J), respectively.</p><p>(K and L) Three-wk-old wild-type (K) and <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> (L) hindlimb skeletons stained with Alcian blue and Alizarin red. The black arrow shows the missing part of proximal femur.</p></div

Depletion of BMP Signaling Causes Interdigital Syndactyly

<div><p>(A–D) Forelimb of adult wild-type (A) and <i>Bmp2^C/C</i>; <i>Prx1::cre</i> mouse (B) and hindlimbs of newborn wild-type (C) mouse and newborn <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> (D) mouse. The black arrow in (B) shows soft tissue syndactyly in <i>Bmp2^C/C</i>; <i>Prx1::cre</i> mouse.</p><p>(E and F) Wild-type and <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i>, respectively, showing acridine orange–stained hindlimbs of E15.5 mouse embryos. Acridine orange stain is in yellow.</p><p>(G and H) Enlarged views of selected regions from (E) and (F), respectively. Black arrow in (G) and (H) show acridine orange–stained apoptotic cells in the interdigital mesenchyme, and asterisk in (H) shows the remnant of the AER.</p><p>(I and J) <i>Fgf8</i> mRNA expression in the hindlimbs of E13.5 wild-type (I) and <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> (J) embryos. The thick black arrows in (J) show Fgf8 mRNA expression.</p></div

In the Absence of BMP2 and BMP4, Osteogenesis Begins During Early Embryonic Development

<div><p>(A–D) Sagittal sections of forelimb from E15.5 embryos. (A and B) Stained with toluidine blue. (A) Distal ulna from wild-type embryo, (B) distal ulna/radius from <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> embryo. Mineralized cartilage is shown by dark purple and osteoid is shown by light blue (red asterisk). (C and D) Humeri sections were hybridized with digoxigenin labeled riboprobe for <i>ColI</i>. (C) Section of wild-type humerus from E15.5 embryo, (D) from <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> E15.5 embryo.</p><p>(E–H) Sagittal sections of zeugopod from E17.5 embryos. (E and F) Stained with toluidine blue while G and H are stained for TRAP (brown stain, red arrow). (E and G) E17.5 proximal radius from wild-type embryo, (F and H) E17.5 proximal ulna/radius from <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> embryo. Although vascularization occurs in the absence of BMP2 and BMP4, few osteoclasts are observed in the mineralized cartilage in <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> mice. Please note all the panels other than (G) and (H) are photographed at ×20, while (G) and (H) were photographed at ×40.</p><p>(I and J) Sections of newborn proximal radius (I) from control animal and newborn proximal ulna/radius (J) from <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> animal were stained with toluidine blue. Red arrow shows the bone marrow cavity in (E) and (I). Note that bone collar (red *) forms at the right time and place in the absence of BMP2 and BMP4. Also note that since <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> mice occasionally have only one bone in zeugopod and the elbow joint is occasionally fused, it is difficult to distinguish between the ulna and the radius.</p></div

Patterning Defects in Limbs Deficient of Different Combinations of BMP Molecules

<div><p>(A–D) <i>Bmp4</i> expression in limb buds from E11.5 wild-type (A and C) and <i>Bmp2^C/C</i>, <i>Bmp7</i> ^{−<i>/</i>−}; <i>Prx1::cre</i> (B and D) mouse embryos. (A and B) Forelimbs, (C and D) hindlimbs.</p><p>(E–H) <i>Shh</i> (E and F) and <i>Fgf8</i> (G and H) expression in E11.5 forelimbs and hindlimbs, respectively. (E and G) Wild-type embryos, (F and H) <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> embryos.</p><p>(I–P) <i>Sox9</i> (I–K) and <i>Msx2</i> (M–O) expression in E12.5 wild-type (I and M) and <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> mouse embryonic forelimbs (J and N) and hindlimbs (K and O). (L and P) <i>Shh</i> expression in E12.5 wild-type (L) and <i>Bmp2^C/C</i>; <i>Bmp4^C/C</i>; <i>Prx1::cre</i> (P) embryonic hindlimbs.</p><p>The red brackets in (F) and (H) show the broadened domains of expressions of <i>Shh</i> and <i>Fgf8</i>, respectively.</p></div

The effect of 25 ng/ml TNFα on proliferation of synovial MSCs.

<p>(A) Experimental design. Synovial MSCs were cultured with 2.5 x 10^-8g/ml TNFα or without TNFα (Control) for 14 days. (B) Representative dishes stained with crystal violet (6 donors). (C) Representative cell colonies stained with crystal violet. Cell number/dish, colony number/dish, and cell number/colony. Average values are shown (n = 6, *p<0.05 by Wilcoxon signed-ranks test).</p