Relationship between the floating toe and the center-of-pressure position in an upright posture among students.

[Purpose] The incidence of floating toes in children is increasing. Although the anteroposterior center of pressure in children is present posteriorly, its relationship with the floating toe is unclear. This study aimed to clarify the relationship between the position of the anteroposterior center of pressure and the floating toe in an upright posture in children. [Participants and Methods] In this cross-sectional study, a Win-Pod (Medicapteurs) platform was used to measure the position of the anteroposterior center of pressure in 208 boys and 195 girls from Japanese elementary schools. Using images of the plantar footprint, floating toes were assessed and the floating toe score was calculated. [Results] The anteroposterior center of pressure position was situated 32.3 ± 8.2% from the heel. The floating toe score of all the participants was 3.5 ± 2.4, with a very high rate of 98%. The floating toe score had a significant, moderate correlation with age, height, weight, and the anteroposterior center-of-pressure position. Multivariate analysis revealed an association between the floating toe score and the anteroposterior center-of-pressure position, height, and weight. [Conclusion] There is significant relationship between the anteroposterior center-of-pressure position and the floating toe score in an upright posture in Japanese elementary school students.

Discovery of SMP-304, a novel benzylpiperidine derivative with serotonin transporter inhibitory activity and 5-HT1A weak partial agonistic activity showing the antidepressant-like effect.

We report the discovery of a novel benzylpiperidine derivative with serotonin transporter (SERT) inhibitory activity and 5-HT1A receptor weak partial agonistic activity showing the antidepressant-like effect. The 3-methoxyphenyl group and the phenethyl group of compound 1, which has weak SERT binding activity, but potent 5-HT1A binding activity, were optimized, leading to compound 35 with potent and balanced dual SERT and 5-HT1A binding activity, but also potent CYP2D6 inhibitory activity. Replacement of the methoxy group in the left part of compound 35 with a larger alkoxy group, such as ethoxy, isopropoxy or methoxy-ethoxy group ameliorated CYP2D6 inhibition, giving SMP-304 as a candidate. SMP-304 with serotonin uptake inhibitory activity and 5-HT1A weak partial agonistic activity, which could work as a 5-HT1A antagonist, displayed faster onset of antidepressant-like effect than a representative SSRI paroxetine in an animal model.

Evaluation of human epididymis protein 4 (HE4) and Risk of Ovarian Malignancy Algorithm (ROMA) as diagnostic tools of type I and type II epithelial ovarian cancer in Japanese women.

Human epididymis protein 4 (HE4) levels and the Risk of Ovarian Malignancy Algorithm (ROMA) have recently been shown to improve the sensitivity and specificity of epithelial ovarian cancer (EOC) diagnosis. We evaluated HE4 levels and ROMA as diagnostic tools of type I and type II EOC in Japanese women. Women who had a pelvic mass on imaging and were scheduled to undergo surgery were enrolled as ovarian mass patients. Serum levels of carbohydrate antigen 125 (CA125) and HE4 were tested in 319 women (131 benign, 19 borderline, 75 malignant, and 94 healthy controls). CA125, HE4, and ROMA were evaluated for sensitivity and by receiver operating characteristics (ROC) in type I and type II EOC. The results showed that, at 75% specificity, the sensitivity of CA125 and HE4 for type II was 92.1% for both markers and for type I was 51.5% and 78.8%, respectively. The sensitivities of ROMA (type I, 84.8% and type II, 97.4%) were better than those of CA125 and HE4. CA125, HE4, and ROMA were all highly accurate markers for type II. For type I, HE4 and ROMA showed better sensitivity than CA125. ROMA displayed the best diagnostic power for type I and type II including for the early stage of type I. In conclusion, HE4, CA125, and ROMA are valuable markers for type II EOC diagnosis. HE4 and ROMA analyses may improve differentiation between type I EOC and a benign mass. Measurement of combined HE4 and CA125 levels provides a more accurate method for EOC diagnosis.

Effects of Competition Level on the Prevalence and Incidence of Lumbar Disk Degeneration in Japanese Collegiate Gymnasts.

Background: Lumbar disk degeneration (LDD) occurs frequently in athletes. Researchers have found that LDD occurs mainly in the lower disks (L4/L5 and L5/S1) in the general and athletic populations. However, a retrospective study showed a high prevalence of LDD in the upper lumbar disks (L1/L2), especially in elite gymnasts. Purpose: To investigate the effect of competition level on the prevalence and incidence of LDD in the upper lumbar disks (L1/L2). Study Design: Cross-sectional study; Level of evidence, 3; and cohort study; Level of evidence, 2. Methods: We conducted 2 studies to evaluate the effect of competition level on the prevalence and incidence of LDD in Japanese collegiate gymnasts. In study 1, a cross-sectional study of 298 collegiate gymnasts was conducted between 2011 and 2015. Competition levels were categorized as regional, national, and international, and T2-weighted magnetic resonance imaging (MRI) was used to evaluate LDD. Chi-square testing was applied to assess differences in the prevalence of LDD and spinal levels among the 3 competition levels. In study 2--a prospective cohort study--LDD progression and its related risk factors were investigated in 51 collegiate gymnasts. Baseline lumbar MRI scans and measurements of physical function (generalized joint laxity and finger-floor distance test) were performed in March 2014. Follow-up lumbar MRI scans were obtained 2 years later, in February 2016. Logistic regression analyses were performed to investigate the relationship between competition level and LDD progression. Results: In study 1, the prevalence of at least 1 degenerated disk in the regional, national, and international groups was 44.2% (19/43), 44.7% (98/219), and 52.8% (19/36), respectively (P = .655). The prevalence of LDD at L1/L2 in the international group was significantly higher than that in the other 2 groups (P = .018). In study 2, the presence of LDD at L1/L2 was associated significantly with international-level competition (adjusted odds ratio, 47.8; 95% CI, 2.75-830.50). Conclusion: In Japanese collegiate gymnasts, competing at the international level was found to be a risk factor for LDD at L1/L2.

Central Tendon Injury Impairs Regional Neuromuscular Activation of the Rectus Femoris Muscle.

We aimed to uncover which rectus femoris strain injury types affect regional activation within the rectus femoris. The rectus femoris has a region-specific functional role; the proximal region of the rectus femoris contributes more than the middle and distal regions during hip flexion. Although a history of strain injury modifies the region-specific functional role within the rectus femoris, it was not obvious which rectus femoris strain injury types affect regional activation within it. We studied 12 soccer players with a history of rectus femoris strain injury. Injury data were obtained from a questionnaire survey and magnetic resonance imaging. To confirm the region-specific functional role of the rectus femoris, surface multichannel electromyographic signals were recorded. Accordingly, eight legs had a history of central tendon injury, four had a history of myofascial junction injury, and four had a healed strain injury. When the injury was limited to the central tendon, the region-specific functional role disappeared. The region-specific functional role was confirmed when the injury was outside the central part. The neuromuscular function was also inhibited when the longitudinal range of the injured region was long. Our findings suggest that a central tendon injury with a long injury length impairs regional neuromuscular activation of the rectus femoris muscle.

Fabrication of GaN/Diamond Heterointerface and Interfacial Chemical Bonding State for Highly Efficient Device Design.

The direct integration of gallium nitride (GaN) and diamond holds much promise for high-power devices. However, it is a big challenge to grow GaN on diamond due to the large lattice and thermal-expansion coefficient mismatch between GaN and diamond. In this work, the fabrication of a GaN/diamond heterointerface is successfully achieved by a surface activated bonding (SAB) method at room temperature. A small compressive stress exists in the GaN/diamond heterointerface, which is significantly smaller than that of the GaN-on-diamond structure with a transition layer formed by crystal growth. A 5.3 nm-thick intermediate layer composed of amorphous carbon and diamond is formed at the as-bonded heterointerface. Ga and N atoms are distributed in the intermediate layer by diffusion during the bonding process. Both the thickness and the sp2 -bonded carbon ratio of the intermediate layer decrease as the annealing temperature increases, which indicates that the amorphous carbon is directly converted into diamond after annealing. The diamond of the intermediate layer acts as a seed crystal. After annealing at 1000 °C, the thickness of the intermediate layer is decreased to approximately 1.5 nm, where lattice fringes of the diamond (220) plane are observed.

[Testicular tumor detected in a male infertility patient with prior orchiopexy].

A 37-year-old man was referred to our hospital with the chief complaint of infertility. Semen analysis at another clinic had demonstrated severe oligozoospermia and he had a history of bilateral orchiopexy for undescended testes. Scrotal ultrasonography revealed a hypoechoic heterogeneous mass (1.5 cm in diameter) within the left testis. With a provisional diagnosis of testicular tumor, left high orchiectomy was performed. Pathological examination of the tumor revealed seminoma. At 12 months after surgery, there has been no evidence of recurrence. Clinicians should be aware of the link between male infertility and testicular cancer, particularly following orchiopexy.

The Effect of a Previous Strain Injury on Regional Neuromuscular Activation Within the Rectus Femoris.

The rectus femoris (RF) has a region-specific functional role; that is, the proximal region of the RF contributes more than the middle and distal regions during hip flexion. This study aimed to investigate whether RF strain injury affected the region-specific functional role of the muscle. We studied seven soccer players with a history of unilateral RF strain injury. Injury data were obtained from a questionnaire survey and magnetic resonance imaging (MRI). Multichannel surface electromyographic (SEMG) signals were recorded from the proximal to distal regions of the RF with 24 electrodes during isometric knee extension and hip flexion. The SEMG signals of each channel during hip flexion were normalised by those during knee extension for the injured and non-injured RF (HF/KE), and compared among the proximal, middle, and distal regions. Six RF strain injuries showed a low signal area in MRI. There was no significant difference in muscle strength between the injured and non-injured RF. While the HF/KE in the proximal region was significantly higher than those in the middle and distal regions in the non-injured RF, a difference in the HF/KE was seen only between the proximal and distal regions of the injured RF. Furthermore, the HF/KE of the most proximal channel in the injured RF was significantly lower than that in the non-injured RF. However, there was no significant difference between injured and non-injured areas in the HF/KE. Our findings suggest that the region-specific functional role of the RF muscle is partly affected by RF strain injury.

Molecular diversity of TEX101, a marker glycoprotein for germ cells monitored with monoclonal antibodies: variety of the molecular characteristics according to subcellular localization within the mouse testis.

TEX101 was characterized as a unique germ cell marker molecule using the specific monoclonal antibody (mAb), TES101. Although this mAb has strong affinity/specificity for TEX101, TES101 mAb loses its reactivity under reducing conditions. In this study, we have generated new mAbs against TEX101 to compensate for the shortcomings of the TES101 mAb using different approaches. First, we immunized mice with the antigen on a baculovirus expression system and isolated new anti-TEX101 mAbs, 6002 and 6035. Second, we raised the mAb Ts4 from spleen cells of an immunologically naive old mouse. Western blot analysis revealed that the new mAbs possess immunoreactivity under reducing/non-reducing conditions. Immunopositive staining of the mAbs against Bouin-fixed sections was observed in spermatocytes, spermatids and testicular spermatozoa, but not in other cells, similar to paraformaldehyde (PFA)-fixed frozen sections stained with TES101 as previously reported. However, whereas the mAbs 6002/6035 mainly showed immunoreactivity only in spermatocytes in PFA-fixed frozen sections, the reactivity of the mAbs to spermatids and testicular spermatozoa was clearly recovered when the PFA-fixed sections were autoclaved or treated with SDS. Peptide mapping and deglycosylation analysis indicated that the epitopes for TES101, 6002 and 6035 are located within TEX101(25-94), whereas Ts4 recognized N-linked carbohydrate moieties on TEX101 in Triton X-100-soluble mouse testicular extracts but not in the extracellular or water-soluble fractions. These results suggest strongly that the molecular association or structure of N-linked carbohydrate moieties of TEX101 varies according to its subcellular localization within the seminiferous tubules. These new mAbs will be valuable tools for further analysis of TEX101, including its function(s).

Association of COL11A1 4603C/T polymorphism with cervical disc degeneration in collegiate wrestlers.

BACKGROUND: The authors previously identified that COL11A1 gene polymorphism is not a susceptibility factor for lumbar disc degeneration (LDD) in athletes. However, the relationship between COL11A1 gene polymorphism and cervical disc degeneration (CDD) remains unclear. We hypothesized that significant associations between COL11A1 4603C/T gene polymorphism and CDD, but not LDD, in collegiate wrestlers exist. This study aims to examine the relationship between CDD, LDD, and COL11A1 4603C/T gene polymorphism in collegiate wrestlers. METHODS: The subjects enrolled in this study were 92 (Study-1) and 123 (Study-2) Japanese collegiate male wrestlers. Study-1 and Study-2 were conducted in 2010-2012 and 2012-2015, respectively. RESULTS: CDD and LDD prevalence among the wrestlers was 51.1% (47/92) and 43.9% (54/123), respectively. We found that COL11A1 4603C/T was significantly associated with CDD, but not with LDD. Using logistic regression analysis with concomitant confounding factors, we further confirmed that COL11A1 4603C/T was a significant risk factor for CDD (co-dominant genetic model [CC vs. CT+TT]: adjusted odds ratio [OR] 2.28; 95% CI: 1.13-4.59; dominant genetic model [CC+CT vs. TT]: adjusted OR 11.71; 95% CI: 1.36-101.06). CONCLUSIONS: Results suggest that COL11A1 4603C/T gene polymorphism is associated with an increased risk of CDD, but not LDD, in Japanese collegiate wrestlers.

In vitro growth and maturation of mouse oocyte-granulosa cell complex from cryopreserved ovaries and achievement of pup birth.

Ovarian tissue banking is a feasible strategy for fertility preservation for young women after cancer treatments. Ovarian tissue, after thawing, is used for several options; orthotopic grafting (normal site), autologous heterotopic grafting and collection of ovarian follicles for culture. Recent reports of live birth encouraged clinicians and researchers to apply this technology to premature ovarian failure (POF) resulting from strong cancer therapy. Grafting, however, carries a risk of malignant cell recurrence. For safety, development of a culture method is necessary but optimum culturing conditions for less-developed follicles abundant in the ovary are not well known. In the present article, the current status of ovarian tissue cryopreservation, and in vitro oocyte growth and maturation from the preserved ovaries are reviewed.

Cervical Intervertebral Disc Degeneration and Low Cervical Extension Independently Associated With a History of Stinger Syndrome.

BACKGROUND: Stinger syndrome frequently occurs in athletes who compete in collision sports. Sharp pain and impairment of neck motion are major symptoms. Cervical intervertebral disc degeneration (CIDD) is also frequently observed in those who compete in collision sports. PURPOSE/HYPOTHESIS: To investigate whether CIDD and neck functionality are related to a history of stinger syndrome. The hypothesis was that a significant relationship exists between CIDD and neck motion and a history of stinger syndrome in Japanese collegiate football players. STUDY DESIGN: Cross-sectional study; Level of evidence, 3. METHODS: A total of 49 male Japanese collegiate football players (mean age, 20.0 ± 1.1 years; mean athletic experience, 3.8 ± 2.3 years; mean height, 172.3 ± 4.8 cm; mean weight, 83.1 ± 12.2 kg) were subdivided into athletes with stinger syndrome (stinger group) and those without (control group). Stinger syndrome was confirmed based on a questionnaire and interview. CIDD was assessed by using T2-weighted magnetic resonance imaging. Range of motion (ROM) and isometric muscle strength were measured for neck function testing. RESULTS: Thirty-nine percent (19/49) of athletes had at least 1 episode of stinger syndrome. The prevalence of CIDD was significantly higher in the stinger group (68%) than in the control group (30%) (P < .01). A statistically significant difference in cervical extension ROM was found between the stinger group (50.9° ± 11.1°) and the control group (60.2° ± 11.4°) (P < .01). Logistic regression analysis showed that CIDD and low cervical extension were independently associated with a history of stinger syndrome. CONCLUSION: Study results suggest that stinger syndrome is associated with CIDD and low cervical extension in collegiate football players.

Upregulation of macrophage migration inhibitory factor and calgizzarin by androgen in TM4 mouse Sertoli cells.

AIM: To identify proteins induced by androgen in Sertoli cells during spermatogenesis. METHODS: We analyzed protein profiles in TM4 Sertoli cells treated with dihydrotestosterone (DHT) using surface enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS). RESULTS: We found increases in the expression of a 5.0-kDa protein at 15 min, an 11.3-kDa protein at 24 h and 4.3 kDa, 5.7 kDa, 5.8 kDa, 9.95 kDa and 9.98 kDa proteins at 48 h after the treatment. In contrast, the expression of 6.3 kDa and 8.6 kDa proteins decreased at 30 min, and 4.9 kDa, 5.0 kDa, 12.4 kDa and 19.8 kDa proteins at 48 h after the treatment. The 11.3-kDa protein was identified as macrophage migration inhibitory factor (MIF) known to having various functions. The 9.98-kDa protein was identified as calgizzarin related to calcium channels. The timing of their expression suggests that MIF and calgizzarin are involved in late regulation of spermatogenesis in Sertoli cells by androgen. CONCLUSION: MIF and calgizzarin are two important androgen-responsive proteins produced by Sertoli cells and they might play a role in regulating spermatogenesis.

Characterization of a novel human sperm-associated antigen 9 (SPAG9) having structural homology with c-Jun N-terminal kinase-interacting protein.

We report a novel SPAG9 (sperm-associated antigen 9) protein having structural homology with JNK (c-Jun N-terminal kinase)-interacting protein 3. SPAG9, a single copy gene mapped to the human chromosome 17q21.33 syntenic with location of mouse chromosome 11, was earlier shown to be expressed exclusively in testis [Shankar, Mohapatra and Suri (1998) Biochem. Biophys. Res. Commun. 243, 561-565]. The SPAG9 amino acid sequence analysis revealed identity with the JNK-binding domain and predicted coiled-coil, leucine zipper and transmembrane domains. The secondary structure analysis predicted an alpha-helical structure for SPAG9 that was confirmed by CD spectra. Microsequencing of higher-order aggregates of recombinant SPAG9 by tandem MS confirmed the amino acid sequence and mono atomic mass of 83.9 kDa. Transient expression of SPAG9 and its deletion mutants revealed that both leucine zipper with extended coiled-coil domains and transmembrane domain of SPAG9 were essential for dimerization and proper localization. Studies of MAPK (mitogenactivated protein kinase) interactions demonstrated that SPAG9 interacted with higher binding affinity to JNK3 and JNK2 compared with JNK1. No interaction was observed with p38alpha or extracellular-signal-regulated kinase pathways. Polyclonal antibodies raised against recombinant SPAG9 recognized native protein in human sperm extracts and localized specifically on the acrosomal compartment of intact human spermatozoa. Acrosome-reacted spermatozoa demonstrated SPAG9 immunofluorescence, indicating its retention on the equatorial segment after the acrosome reaction. Further, anti-SPAG9 antibodies inhibited the binding of human spermatozoa to intact human oocytes as well as to matched hemizona. This is the first report of sperm-associated JNK-binding protein that may have a role in spermatozoa-egg interaction.

Asymmetrical and smaller size of trunk muscles in combat sports athletes with lumbar intervertebral disc degeneration.

BACKGROUND: Lumbar intervertebral disc degeneration (LDD) frequently occurs in athletes. Associations between LDD and trunk muscles still remain unclear. PURPOSE: This study examined whether there is an association between the prevalence of LDD and the symmetry and size of the cross-sectional areas (CSAs) of the trunk muscles in combat sports athletes. METHODS: Participants in this study were 151 collegiate male combat sports athletes. A total of 755 lumbar intervertebral discs from L1-2 to L5-S1 in 151 athletes were assessed using magnetic resonance imaging (MRI) and a comprehensive grading system of LDD (grades I-V). All 151 athletes were divided into 2 groups: LDD and non-LDD. CSAs of trunk muscles at the L3-4 disc level were measured using MRI. RESULTS: Sixty-nine athletes had LDD at 1 or more disc levels (45.7 %). The LDD grade for the lower 2 disc levels was significantly higher than that for the other disc levels (p < 0.001). The CSAs of the left and right sides in trunk muscles were significantly asymmetrical, independent of the LDD which was prevalent in the disc levels (obliques: p = 0.040; quadratus lumborum: p < 0.001). The relative CSAs of trunk muscles to their body weight in the LDD group were significantly smaller than those in the non-LDD group (rectus abdominis: p = 0.011; obliques: p = 0.024; quadratus lumborum: p = 0.006; lumbar erector spinae plus multifidus: p = 0.001). CONCLUSION: This study suggests that the prevalence of LDD is associated with asymmetrical and relatively smaller CSAs of trunk muscles in combat sports athletes.

Immunogenicity and contraceptive potential of recombinant human sperm associated antigen (SPAG9).

Human sperm-associated antigen 9 (hSPAG9) is a potential target for sperm-based contraceptive vaccine in lieu of its location on the sperm acrosomal compartment and its implication in sperm-egg interaction. SPAG9 is an acrosomal molecule which is not only restricted to a specific region (domain) of the acrosome but also undergoes relocation to the equatorial region in a stage-specific manner during acrosome reaction, demonstrating its potential role in sperm-egg binding. Human SPAG9 nucleotide sequence revealed 94% identity with macaque SPAG9 and 96.8% with baboon SPAG9 over the entire sequence. The amino acid sequence comparison of human SPAG9 with macaque and baboon revealed an overall homology of 84.9% and 90.6%, respectively. The presence of a high level of homology at the amino acid and nucleotide levels indicates that SPAG9 is conserved in macaque, baboon and human sharing common function and common origin in the biological past. Immunogenicity studies were carried in rats, which demonstrated that recombinant hSPAG9 protein adsorbed on alum is highly immunogenic. Antibodies thus generated after immunization reacted with recombinant human SPAG9 (rhSPAG9) and native SPAG9 protein from human sperm in Western blot analysis. In an in vitro assay, anti-rhSPAG9 antibodies inhibited sperm adherence to or penetration in zona-free hamster egg penetration test. Further, anti-SPAG9 antibodies inhibited the binding of human sperm to intact human oocyte as well as to matched hemi-zonae, indicating that the recombinant protein is a suitable contraceptive vaccinogen. Together these results demonstrate that the rhSPAG9 adsorbed on alum is immunogenic in nature, which is a permissible adjuvant for immunogenicity and fertility trials in non-human primates.

Target antigens for sperm-immobilizing antibodies found in infertile women.

Complement-dependent sperm-immobilizing antibodies (SI-Abs) have frequently been detected in the sera of infertile women. Analyses using monoclonal antibodies as well as patients' antisperm antibodies have shown that the carbohydrate moieties of sperm and seminal plasma are major epitopes of these antibodies. In the present study, we characterized the antigen molecules recognized by human monoclonal SI-Ab (MAb H6-3C4) derived from a patient with strong SI-Ab activity. Immunofluorescent staining showed that MAb H6-3C4 reacted with ejaculated human sperm and the epididymis but not with the testes and other somatic organs including the spleen. In Western blot analysis, MAb H6-3C4 reacted to antigen molecules of Mr 15-27 kDa present in the methanol-chloroform extracts of either ejaculated sperm or seminal plasma. Amino acid sequence analysis of the reactive molecule revealed that the amino acid sequence was identical to the core peptide of CD52, a GPI-anchored protein on lymphocytes. Furthermore, the epitope for MAb H6-3C4 was found to be the specific N-linked carbohydrate for the male reproductive tract. These results suggest that SI-Abs in some infertile women target a sperm-specific carbohydrate antigen on CD52 molecules which are secreted from the epididymis and coat the sperm surface.

Follicular dysfunction induced by autoimmunity to zona pellucida.

The mammalian zona pellucida is an extracellular matrix that occurs in growing oocytes, ovulated eggs and pre-implantation embryos, and is known to be involved in several important events during ovarian folliculogenesis and fertilization. Since the zona pellucida is formed at an early stage of oocyte growth, circulating antibodies against zona pellucida may impair ovarian function. In this article we discuss whether anti-zona antibodies cause ovarian dysfunction and infertility. The discussion is based on clinical examination and animal experiments including the following approaches: 1/ immunological method using solubilized human zona pellucida detected anti-zona antibody with a high frequency in infertile patients, especially premature ovarian failure syndrome; 2/ in vivo experiment using hamsters showed that some, but not all, animals experienced ovarian failure after immunization with hamster recombinant zona proteins; 3/ in vitro experiment using mouse isolated ovarian follicles showed significant inhibitory effects on follicular growth and oocyte development. We concluded that anti-zona antibody may be involved in causing ovarian failure.

Layer matching epitaxy of NiO thin films on atomically stepped sapphire (0001) substrates.

Thin-film epitaxy is critical for investigating the original properties of materials. To obtain epitaxial films, careful consideration of the external conditions, i.e. single-crystal substrate, temperature, deposition pressure and fabrication method, is significantly important. In particular, selection of the single-crystal substrate is the first step towards fabrication of a high-quality film. Sapphire (single-crystalline α-Al2O3) is commonly used in industry as a thin-film crystal-growth substrate, and functional thin-film materials deposited on sapphire substrates have found industrial applications. However, while sapphire is a single crystal, two types of atomic planes exist in accordance with step height. Here we discuss the need to consider the lattice mismatch for each of the sapphire atomic layers. Furthermore, through cross-sectional transmission electron microscopy analysis, we demonstrate the uniepitaxial growth of cubic crystalline thin films on bistepped sapphire (0001) substrates.