Cotton(Gossypium hirsutum L.) fibers,one of the most important natural raw materials for the textile industry,are highly elongated trichomes from epidermal cells of cotton ovules.Among the longest plant cells ever cha...Cotton(Gossypium hirsutum L.) fibers,one of the most important natural raw materials for the textile industry,are highly elongated trichomes from epidermal cells of cotton ovules.Among the longest plant cells ever characterized,cotton fiber is an ideal system for studying plant cell elongation.展开更多
In this project,we aim to elucidate the molecular mechanism controlling initiation and elongation of tetraploid Gossypium hirsutum fiber cells by setting up a high throughput custom-designed
The regulatory mechanism that controls the sustained cotton fiber cell elongation is gradually being elucidated by coupling genome-wide transcriptome profiling with systematic biochemical and physiological studies.Ver...The regulatory mechanism that controls the sustained cotton fiber cell elongation is gradually being elucidated by coupling genome-wide transcriptome profiling with systematic biochemical and physiological studies.Very long chain fatty acids(VLCFA),H2O2,and several types of plant展开更多
A comparative proteomic analysis was applied to explore the mechanism of fiber cell development in cotton.Initially,an efficient protein preparation method was established for proteomic analysis
AIM:To investigate peripapillary retinal nerve fiber layer (RNFL), macula and ganglion cell layer thicknesses (GCC) in amblyopic eyes with spectral domain optical coherence tomography (SD-OCT). METHODS:Thirty six pati...AIM:To investigate peripapillary retinal nerve fiber layer (RNFL), macula and ganglion cell layer thicknesses (GCC) in amblyopic eyes with spectral domain optical coherence tomography (SD-OCT). METHODS:Thirty six patients with a history of unilateral amblyopia and thirty two children who had emmetropia without amblyopia were included in this study. In this institutional study, 36 eyes of 36 patients with amblyopia (AE), 36 fellow eyes without amblyopia (FE), and 32 eyes of 32 normal subjects (NE) were included. RNFL, GCC and macular thickness measurements were performed with RS-3000 OCT Retina Scan (Nidek Inc CA. USA). RESULTS:The mean global thicknesses of the RNFL were 113.22 ±21.47, 111.57 ±18.25, 109.96 ±11.31μm in the AE, FE, and NE, respectively. There was no statistically significant difference for mean global RNFL thickness among the eyes (P =0.13). The mean thicknesses of the macula were 258.25±18.31, 258.75±19.54, 248.62±10.57μm in the AE, FE and NE, respectively. There was no statistically significant difference for thickness of macula among the eyes (P =0.06). The GCC was investigated into two parts:superior and inferior. The mean thicknesses of superior GCC were 102.57 ±13.32, 103.32 ±10.64, 100.52 ± 5.88μm in the AE, FE, and NE, respectively. The mean thicknesses of inferior GCC were 103.82 ±12.60, 107.82 ± 12.33, 105.86±10.79μm in the AE, FE and NE, respectively. There was no statistically significant difference for thickness of superior and inferior GCC between the eyes (P =0.63, P =0.46). ·CONCLUSION:The macular thicknesses of AE and FE were greater than the NE, although it was not statistically significant. Amblyopia does not seem to have a profound effect on the RNFL, macula and GCC.展开更多
Calotropis procera, commonly known as "milkweed", possesses long seed trichomes for seed dispersal and has the ability to survive under harsh conditions such as drought and salinity. Aquaporins are water cha...Calotropis procera, commonly known as "milkweed", possesses long seed trichomes for seed dispersal and has the ability to survive under harsh conditions such as drought and salinity. Aquaporins are water channel proteins expressed in all land plants, divided into five subfamilies plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), NOD26-like proteins (NIPs), small basic intrinsic proteins (SIPs), and the unfamiliar X intrinsic proteins (XIPs). PIPs constitute the largest group of water channel proteins that are involved in different developmental and regulatory mechanisms including water permeability, cell elongation, and stomata opening. Aquaporins are also involved in abiotic stress tolerance and cell expansion mechanisms, but their role in seed trichomes (fiber cells) has never been investigated. A large number of clones isolated from C. procera fiber cDNA library showed sequence homology to PIPs. Both expressed sequence tags (ESTs) and real-time polymerase chain reaction (PCR) studies revealed that the transcript abundance of this gene family in fiber cells of C. procera is greater than that of cotton. Full-length cDNAs of CpPIP1 and CpPIP2 were isolated from C. procera fiber cDNA library and used for constructing plant expression vectors under constitutive (2×35S) and trichome-specific (GhLTP3) promoters. Transgenic tobacco plants were developed via Agrobacterium-mediated transformation. The phenotypic characteristics of the plants were observed after confirming the integration of transgene in plants. It was observed that CpPIP2 expression cassette under 2×35S and GhLTP3 promoter enhanced the numbers of stem and leave trichomes. However, 2×35S::CpPIP2 has a more amplified effect on trichome density and length than GhLTP3::CpPIP2 and other PIP constructs. These findings imply the role of C. procera PIP aquaporins in fiber cell elongation. The PIPs-derived cell expansion mechanism may be exploited through transgenic approaches for improvement of fiber staple length in cotton and boosting of defense against sucking insects by enhancing plant pubescence.展开更多
AIM:To evaluate the structural injure patterns in peripapillary retinal fiber layer (pRNFL), retinal ganglion cell layer (RGCL) and their correlations to visual function in various mitochondrial optic neuropathi...AIM:To evaluate the structural injure patterns in peripapillary retinal fiber layer (pRNFL), retinal ganglion cell layer (RGCL) and their correlations to visual function in various mitochondrial optic neuropathies (MON) to offer help to their differential diagnosis.METHODS:Totally 32 MON patients (60 eyes) were recruited within 6mo after clinical onsets, including 20 Leber hereditary optic neuropathy (LHON) patients (37eyes), 12 ethambutol-induced optic neuropathy (EON)patients (23 eyes), and 41 age-gender matched healthy controls (HC, 82 eyes). All subjects had pRNFL and RGCL examinations with optic coherence tomography (OCT) and visual function tests.RESULTS:In the early stages of MON, the temporal pRNFL thickness decreased (66.09±22.57μm), but increased in other quadrants, compared to HC (76.95±14.81μm). The other quadrants remaining stable for LHON and EON patients besides the second hour sector of pRNFL thickness reduced and the temporal pRNFL decreased (56.78±15.87μm) for EON. Total macular thickness in MON reduced remarkably(279.25±18.90μm; P=0.015), which mainly occurring in the inner circle (3 mm diameter of circle) and the nasal temporal sectors in the outer circle (5.5 mm diameter of circle), in contrast to those in HC. RGCL thickness reduced in each sector of the macula (61.90±8.73μm; P≤0.001). It strongly showed the correlationship of best corrected visual acuity (R=0.50, P=0.0003) and visual field injury (R=0.54,P=0.0002) in MON patients.CONCLUSION:OCT is a potential tool for detecting structural alterations in the optic nerves of various MON. Different types of MON may have different damage patterns.展开更多
Abstract: To enable the use of metallic components in direct methanol fuel cells (DMFCs), issues related to corrosion resistance must be considered because of an acid environment induced by the solid electrolyte. I...Abstract: To enable the use of metallic components in direct methanol fuel cells (DMFCs), issues related to corrosion resistance must be considered because of an acid environment induced by the solid electrolyte. In this study, we report the electrochemical behaviors of metal-fiber-based porous sintered components in a simulated corrosive environment of DMFCs. Three materials were evaluated: pure copper, AISI304, and AISI316L. The environmental factors and related mechanisms affecting the corrosion behaviors were analyzed. The results demonstrated that AISI316L exhibits the best performance. A higher SO4^2- concentration increases the risk of material corrosion, whereas an increase in methanol concentration inhibits corrosion. The morphological features of the corroded samples were also characterized in this study.展开更多
The work compared the differences of low fiber mutation rice (LF, Nendao) that selectedthrough gamma-ray (γ) with parental variety Shuangkezao (CK) on their biologicaldevelopment and cell wall composing after rice he...The work compared the differences of low fiber mutation rice (LF, Nendao) that selectedthrough gamma-ray (γ) with parental variety Shuangkezao (CK) on their biologicaldevelopment and cell wall composing after rice heading stage. Comparing with parentalrice, LF rice revealed an advantage on its vegetative growing by increasing the yieldsof leave blade, leave sheath and stem for 27.77, 30.19 and 37.96% respectively. And thecellulose content of LF rice straw was decreased remarkably for 23.9%, the hemicellulose,lignin and biogenic silicon contents were increased contrarily for 11.94, 8.79 and 5.60%respectively. Moreover, the crude protein content was increased by 20.71% for LF rice andwith an improvement on its solubility for 63.49% concomitantly. The results indicatedthat the low-fiber mutation rice exhibited its potential as a fodder-rice variety or asdual-purpose rice to improve fiber degradability of straw.展开更多
AIM: To determine the thickness of the retinal ganglion cell-inner plexiform layer(GCIPL) and the retinal nerve fiber layer(RNFL) in patients with neuromyelitis optica(NMO).METHODS: We conducted a cross-sectio...AIM: To determine the thickness of the retinal ganglion cell-inner plexiform layer(GCIPL) and the retinal nerve fiber layer(RNFL) in patients with neuromyelitis optica(NMO).METHODS: We conducted a cross-sectional study that included 30 NMO patients with a total of 60 eyes. Based on the presence or absence of optic neuritis(ON), subjects were divided into either the NMO-ON group(30 eyes) or the NMO-ON contra group(10 eyes). A detailed ophthalmologic examination was performed for each group; subsequently, the GCIPL and the RNFL were measured using highdefinition optical coherence tomography(OCT). RESULTS: In the NMO-ON group, the mean GCIPL thickness was 69.28±21.12 μm, the minimum GCIPL thickness was 66.02±10.02 μm, and the RNFL thickness were 109.33±11.23, 110.47±3.10, 64.92±12.71 and 71.21±50.22 μm in the superior, inferior, temporal and nasal quadrants, respectively. In the NMO-ON contra group, the mean GCIPL thickness was 85.12±17.09 μm, the minimum GCIPL thickness was 25.39±25.1 μm, and the RNFL thicknesses were 148.33±23.22, 126.36±23.45, 82.21±22.30 and 83.36±31.28 μm in the superior, inferior, temporal and nasal quadrants, respectively. In the control group, the mean GCIPL thickness was 86.98±22.37 μm, the minimum GCIPL thickness was 85.28±10.75 μm, and the RNFL thicknesses were 150.22±22.73, 154.79±60.23, 82.33±7.01 and 85.62±13.81 μm in the superior, inferior, temporal and nasal quadrants, respectively. The GCIPL and RNFL were thinner in the NMO-ON contra group than in the control group(P〈0.05); additionally, the RNFL was thinner in the inferior quadrant in the NMO-ON group than in the control group(P〈0.05). Significant correlations were observed between the GCIPL and RNFL thickness measurements as well as between thickness measurements and the two visual field parameters of mean deviation(MD) and corrected pattern standard deviation(PSD) in the NMO-ON group(P〈0.05). CONCLUSION: The thickness of the GCIPL and RNFL, as measured using OCT, may indicate optic nerve damage in patients with NMO.展开更多
AIM:To evaluate retinal nerve fiber layer(RNFL)thickness analysis of peripapillary optic nerve head(PONH) and macula as well as ganglion cell-inner plexiform layer(GCIPL) thickness in obese children.· METH...AIM:To evaluate retinal nerve fiber layer(RNFL)thickness analysis of peripapillary optic nerve head(PONH) and macula as well as ganglion cell-inner plexiform layer(GCIPL) thickness in obese children.· METHODS:Eighty-five children with obesity and 30 controls were included in the study.The thicknesses of the PONH and macula of each subject's right eye were measured by high-resolution spectral-domain optic coherence tomography(OCT).· RESULTS:The RNFL thicknesses of central macular and PONH were similar between the groups(all P 〉0.05).The GCIPL thickness was also similar between the groups.However,the RNFL thickness of temporal outer macula were 261.7±13.7 and 268.9±14.3 μm for the obesity and the control group,respectively(P =0.034).· CONCLUSION:Obesity may cause a reduction in temporal outer macular RNFL thickness.展开更多
The present study was undertaken to determine the variability in fiber cell morphology and its length among wood species of the Tamaulipan Thorn Scrub, Northeastern Mexico, used for various uses such as timber for fur...The present study was undertaken to determine the variability in fiber cell morphology and its length among wood species of the Tamaulipan Thorn Scrub, Northeastern Mexico, used for various uses such as timber for furniture, fence, post, firewood and sources of forage. The results reveal large variability in fiber cell morphology and fiber cell dimensions. The species are classified on the basis of its fiber cell morphology and fiber cell length and recommended for their possible utilization for different purposes. Wood having fibre cells with broad lumen and thin wall could be suitable for the manufacture of paper documented in the literature viz. Acacia farnesciana, Caesalpinia mexicana, Cordia boisiieri, Diospyros palmeri, Forestieria angustifolia, Morus celtifolia, Prosopis laevigata. This technique can be used in preliminary screening of woody species for its utilization for different purposes. Further studies are needed to confirm.展开更多
文摘Cotton(Gossypium hirsutum L.) fibers,one of the most important natural raw materials for the textile industry,are highly elongated trichomes from epidermal cells of cotton ovules.Among the longest plant cells ever characterized,cotton fiber is an ideal system for studying plant cell elongation.
文摘In this project,we aim to elucidate the molecular mechanism controlling initiation and elongation of tetraploid Gossypium hirsutum fiber cells by setting up a high throughput custom-designed
文摘The regulatory mechanism that controls the sustained cotton fiber cell elongation is gradually being elucidated by coupling genome-wide transcriptome profiling with systematic biochemical and physiological studies.Very long chain fatty acids(VLCFA),H2O2,and several types of plant
文摘A comparative proteomic analysis was applied to explore the mechanism of fiber cell development in cotton.Initially,an efficient protein preparation method was established for proteomic analysis
文摘AIM:To investigate peripapillary retinal nerve fiber layer (RNFL), macula and ganglion cell layer thicknesses (GCC) in amblyopic eyes with spectral domain optical coherence tomography (SD-OCT). METHODS:Thirty six patients with a history of unilateral amblyopia and thirty two children who had emmetropia without amblyopia were included in this study. In this institutional study, 36 eyes of 36 patients with amblyopia (AE), 36 fellow eyes without amblyopia (FE), and 32 eyes of 32 normal subjects (NE) were included. RNFL, GCC and macular thickness measurements were performed with RS-3000 OCT Retina Scan (Nidek Inc CA. USA). RESULTS:The mean global thicknesses of the RNFL were 113.22 ±21.47, 111.57 ±18.25, 109.96 ±11.31μm in the AE, FE, and NE, respectively. There was no statistically significant difference for mean global RNFL thickness among the eyes (P =0.13). The mean thicknesses of the macula were 258.25±18.31, 258.75±19.54, 248.62±10.57μm in the AE, FE and NE, respectively. There was no statistically significant difference for thickness of macula among the eyes (P =0.06). The GCC was investigated into two parts:superior and inferior. The mean thicknesses of superior GCC were 102.57 ±13.32, 103.32 ±10.64, 100.52 ± 5.88μm in the AE, FE, and NE, respectively. The mean thicknesses of inferior GCC were 103.82 ±12.60, 107.82 ± 12.33, 105.86±10.79μm in the AE, FE and NE, respectively. There was no statistically significant difference for thickness of superior and inferior GCC between the eyes (P =0.63, P =0.46). ·CONCLUSION:The macular thicknesses of AE and FE were greater than the NE, although it was not statistically significant. Amblyopia does not seem to have a profound effect on the RNFL, macula and GCC.
文摘Calotropis procera, commonly known as "milkweed", possesses long seed trichomes for seed dispersal and has the ability to survive under harsh conditions such as drought and salinity. Aquaporins are water channel proteins expressed in all land plants, divided into five subfamilies plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), NOD26-like proteins (NIPs), small basic intrinsic proteins (SIPs), and the unfamiliar X intrinsic proteins (XIPs). PIPs constitute the largest group of water channel proteins that are involved in different developmental and regulatory mechanisms including water permeability, cell elongation, and stomata opening. Aquaporins are also involved in abiotic stress tolerance and cell expansion mechanisms, but their role in seed trichomes (fiber cells) has never been investigated. A large number of clones isolated from C. procera fiber cDNA library showed sequence homology to PIPs. Both expressed sequence tags (ESTs) and real-time polymerase chain reaction (PCR) studies revealed that the transcript abundance of this gene family in fiber cells of C. procera is greater than that of cotton. Full-length cDNAs of CpPIP1 and CpPIP2 were isolated from C. procera fiber cDNA library and used for constructing plant expression vectors under constitutive (2×35S) and trichome-specific (GhLTP3) promoters. Transgenic tobacco plants were developed via Agrobacterium-mediated transformation. The phenotypic characteristics of the plants were observed after confirming the integration of transgene in plants. It was observed that CpPIP2 expression cassette under 2×35S and GhLTP3 promoter enhanced the numbers of stem and leave trichomes. However, 2×35S::CpPIP2 has a more amplified effect on trichome density and length than GhLTP3::CpPIP2 and other PIP constructs. These findings imply the role of C. procera PIP aquaporins in fiber cell elongation. The PIPs-derived cell expansion mechanism may be exploited through transgenic approaches for improvement of fiber staple length in cotton and boosting of defense against sucking insects by enhancing plant pubescence.
基金Supported by the National High Technology Research and Development Program of China(863 Program,No.2015AA020511)
文摘AIM:To evaluate the structural injure patterns in peripapillary retinal fiber layer (pRNFL), retinal ganglion cell layer (RGCL) and their correlations to visual function in various mitochondrial optic neuropathies (MON) to offer help to their differential diagnosis.METHODS:Totally 32 MON patients (60 eyes) were recruited within 6mo after clinical onsets, including 20 Leber hereditary optic neuropathy (LHON) patients (37eyes), 12 ethambutol-induced optic neuropathy (EON)patients (23 eyes), and 41 age-gender matched healthy controls (HC, 82 eyes). All subjects had pRNFL and RGCL examinations with optic coherence tomography (OCT) and visual function tests.RESULTS:In the early stages of MON, the temporal pRNFL thickness decreased (66.09±22.57μm), but increased in other quadrants, compared to HC (76.95±14.81μm). The other quadrants remaining stable for LHON and EON patients besides the second hour sector of pRNFL thickness reduced and the temporal pRNFL decreased (56.78±15.87μm) for EON. Total macular thickness in MON reduced remarkably(279.25±18.90μm; P=0.015), which mainly occurring in the inner circle (3 mm diameter of circle) and the nasal temporal sectors in the outer circle (5.5 mm diameter of circle), in contrast to those in HC. RGCL thickness reduced in each sector of the macula (61.90±8.73μm; P≤0.001). It strongly showed the correlationship of best corrected visual acuity (R=0.50, P=0.0003) and visual field injury (R=0.54,P=0.0002) in MON patients.CONCLUSION:OCT is a potential tool for detecting structural alterations in the optic nerves of various MON. Different types of MON may have different damage patterns.
基金financially supported by the Natural Science Foundation of Guangdong Province, China (No. S2013040016899)the Fundamental Research Funds for Central Universities of China (No. 2013ZM0003)+1 种基金the National Natural Science Foundation of China (No. 51275180)the Open Fund of Shanghai Key Laboratory of Digital Manufacture for Thin-walled Structures (No. 2013001)
文摘Abstract: To enable the use of metallic components in direct methanol fuel cells (DMFCs), issues related to corrosion resistance must be considered because of an acid environment induced by the solid electrolyte. In this study, we report the electrochemical behaviors of metal-fiber-based porous sintered components in a simulated corrosive environment of DMFCs. Three materials were evaluated: pure copper, AISI304, and AISI316L. The environmental factors and related mechanisms affecting the corrosion behaviors were analyzed. The results demonstrated that AISI316L exhibits the best performance. A higher SO4^2- concentration increases the risk of material corrosion, whereas an increase in methanol concentration inhibits corrosion. The morphological features of the corroded samples were also characterized in this study.
文摘The work compared the differences of low fiber mutation rice (LF, Nendao) that selectedthrough gamma-ray (γ) with parental variety Shuangkezao (CK) on their biologicaldevelopment and cell wall composing after rice heading stage. Comparing with parentalrice, LF rice revealed an advantage on its vegetative growing by increasing the yieldsof leave blade, leave sheath and stem for 27.77, 30.19 and 37.96% respectively. And thecellulose content of LF rice straw was decreased remarkably for 23.9%, the hemicellulose,lignin and biogenic silicon contents were increased contrarily for 11.94, 8.79 and 5.60%respectively. Moreover, the crude protein content was increased by 20.71% for LF rice andwith an improvement on its solubility for 63.49% concomitantly. The results indicatedthat the low-fiber mutation rice exhibited its potential as a fodder-rice variety or asdual-purpose rice to improve fiber degradability of straw.
基金Supported by Science and Technology Bureau Project Fund of Wenzhou, China (No.Y20160460)
文摘AIM: To determine the thickness of the retinal ganglion cell-inner plexiform layer(GCIPL) and the retinal nerve fiber layer(RNFL) in patients with neuromyelitis optica(NMO).METHODS: We conducted a cross-sectional study that included 30 NMO patients with a total of 60 eyes. Based on the presence or absence of optic neuritis(ON), subjects were divided into either the NMO-ON group(30 eyes) or the NMO-ON contra group(10 eyes). A detailed ophthalmologic examination was performed for each group; subsequently, the GCIPL and the RNFL were measured using highdefinition optical coherence tomography(OCT). RESULTS: In the NMO-ON group, the mean GCIPL thickness was 69.28±21.12 μm, the minimum GCIPL thickness was 66.02±10.02 μm, and the RNFL thickness were 109.33±11.23, 110.47±3.10, 64.92±12.71 and 71.21±50.22 μm in the superior, inferior, temporal and nasal quadrants, respectively. In the NMO-ON contra group, the mean GCIPL thickness was 85.12±17.09 μm, the minimum GCIPL thickness was 25.39±25.1 μm, and the RNFL thicknesses were 148.33±23.22, 126.36±23.45, 82.21±22.30 and 83.36±31.28 μm in the superior, inferior, temporal and nasal quadrants, respectively. In the control group, the mean GCIPL thickness was 86.98±22.37 μm, the minimum GCIPL thickness was 85.28±10.75 μm, and the RNFL thicknesses were 150.22±22.73, 154.79±60.23, 82.33±7.01 and 85.62±13.81 μm in the superior, inferior, temporal and nasal quadrants, respectively. The GCIPL and RNFL were thinner in the NMO-ON contra group than in the control group(P〈0.05); additionally, the RNFL was thinner in the inferior quadrant in the NMO-ON group than in the control group(P〈0.05). Significant correlations were observed between the GCIPL and RNFL thickness measurements as well as between thickness measurements and the two visual field parameters of mean deviation(MD) and corrected pattern standard deviation(PSD) in the NMO-ON group(P〈0.05). CONCLUSION: The thickness of the GCIPL and RNFL, as measured using OCT, may indicate optic nerve damage in patients with NMO.
基金partially presented at the 7~(th) Mediteretina Club International Meeting,April 17-20,2014,Istanbul
文摘AIM:To evaluate retinal nerve fiber layer(RNFL)thickness analysis of peripapillary optic nerve head(PONH) and macula as well as ganglion cell-inner plexiform layer(GCIPL) thickness in obese children.· METHODS:Eighty-five children with obesity and 30 controls were included in the study.The thicknesses of the PONH and macula of each subject's right eye were measured by high-resolution spectral-domain optic coherence tomography(OCT).· RESULTS:The RNFL thicknesses of central macular and PONH were similar between the groups(all P 〉0.05).The GCIPL thickness was also similar between the groups.However,the RNFL thickness of temporal outer macula were 261.7±13.7 and 268.9±14.3 μm for the obesity and the control group,respectively(P =0.034).· CONCLUSION:Obesity may cause a reduction in temporal outer macular RNFL thickness.
文摘The present study was undertaken to determine the variability in fiber cell morphology and its length among wood species of the Tamaulipan Thorn Scrub, Northeastern Mexico, used for various uses such as timber for furniture, fence, post, firewood and sources of forage. The results reveal large variability in fiber cell morphology and fiber cell dimensions. The species are classified on the basis of its fiber cell morphology and fiber cell length and recommended for their possible utilization for different purposes. Wood having fibre cells with broad lumen and thin wall could be suitable for the manufacture of paper documented in the literature viz. Acacia farnesciana, Caesalpinia mexicana, Cordia boisiieri, Diospyros palmeri, Forestieria angustifolia, Morus celtifolia, Prosopis laevigata. This technique can be used in preliminary screening of woody species for its utilization for different purposes. Further studies are needed to confirm.