Adaptive Planar Chirality of Pillar[5]arenes Invertible by n-Alkane Lengths.

Chirality of host molecules can be induced and/or inverted by the guest molecules. However, the adapting chirality of hosts to the length of n-alkanes remains a great challenge because n-alkanes are neutral, achiral, and linear molecules, resulting in a weak interaction with most compounds. Herein, we report a system with chirality adapted to n-alkane lengths, using a pillar[5]arene-based macrocyclic host, S-Br, which contains five stereogenic carbons and five terminal bromine atoms on each rim. The electron-rich cavity of S-Br could include n-alkanes and the planar-chiral isomers sensitively inverted in response to the lengths of the complexed n-alkanes. The inclusion of a short n-alkane such as n-pentane made S-Br more inclined to be in the pS-form, whereas the inclusion of long n-alkanes such as n-heptane made the pR-form more favorable. The difference in the stability of the isomers was supported by the crystal structures and the theoretical calculations. Furthermore, temperature drives the adaptive chirality of S-Br with n-alkanes. An n-alkane with middle length, n-hexane, showed the dominance of the pR-form of S-Br at a higher temperature, whereas the pS-form was shown at a lower temperature.

Real-time chirality transfer monitoring from statistically random to discrete homochiral nanotubes.

Real time monitoring of chirality transfer processes is necessary to better understand their kinetic properties. Herein, we monitor an ideal chirality transfer process from a statistically random distribution to a diastereomerically pure assembly in real time. The chirality transfer is based on discrete trimeric tubular assemblies of planar chiral pillar[5]arenes, achieving the construction of diastereomerically pure trimers of pillar[5]arenes through synergistic effect of ion pairing between a racemic rim-differentiated pillar[5]arene pentaacid bearing five benzoic acids on one rim and five alkyl chains on the other, and an optically resolved pillar[5]arene decaamine bearing ten amines. When the decaamine is mixed with the pentaacid, the decaamine is sandwiched by two pentaacids through ten ion pairs, initially producing a statistically random mixture of a homochiral trimer and two heterochiral trimers. The heterochiral trimers gradually dissociate and reassemble into the homochiral trimers after unit flipping of the pentaacid, leading to chirality transfer from the decaamine and producing diastereomerically pure trimers.

Estimation of breeding population size using DNA-based pedigree reconstruction in brown bears.

Robust estimates of demographic parameters are critical for effective wildlife conservation and management but are difficult to obtain for elusive species. We estimated the breeding and adult population sizes, as well as the minimum population size, in a high-density brown bear population on the Shiretoko Peninsula, in Hokkaido, Japan, using DNA-based pedigree reconstruction. A total of 1288 individuals, collected in and around the Shiretoko Peninsula between 1998 and 2020, were genotyped at 21 microsatellite loci. Among them, 499 individuals were identified by intensive genetic sampling conducted in two consecutive years (2019 and 2020) mainly by noninvasive methods (e.g., hair and fecal DNA). Among them, both parents were assigned for 330 bears, and either maternity or paternity was assigned to 47 and 76 individuals, respectively. The subsequent pedigree reconstruction indicated a range of breeding and adult (≥4 years old) population sizes: 128-173 for female breeders and 66-91 male breeders, and 155-200 for female adults and 84-109 male adults. The minimum population size was estimated to be 449 (252 females and 197 males) in 2019. Long-term continuous genetic sampling prior to a short-term intensive survey would enable parentage to be identified in a population with a high probability, thus enabling reliable estimates of breeding population size for elusive species.

Diel and monthly activity pattern of brown bears and sika deer in the Shiretoko Peninsula, Hokkaido, Japan.

Mammals exhibit several types of diel activity pattern, including nocturnal, diurnal, crepuscular, and cathemeral. These patterns vary inter- and intra-specifically and are affected by environmental factors, individual status, and interactions with other individuals or species. Determining the factors that shape diel activity patterns is challenging but essential for understanding the behavioral ecology of animal species, and for wildlife conservation and management. Using camera-trap surveys, we investigated the species distributions and activity patterns of terrestrial mammals on the Shiretoko Peninsula, Hokkaido, Japan, with particular focus on brown bears and sika deer. From June to October 2019, a total of 7,530 observations were recorded by 65 camera-traps for eight species, including two alien species. The diel activity pattern of brown bears was diurnal/crepuscular, similar to that of bears in North America, but different from European populations. Bear observations were more frequent during the autumnal hyperphagia period, and adult females and sub-adults were more diurnal than adult males. In addition, bears inside the protected area were more diurnal than those outside it. These findings suggest that appetite motivation, competitive interactions between conspecifics, and human activities potentially affect bear activity patterns. Similar to other sika deer populations and other deer species, the diel activity patterns of sika deer were crepuscular. Deer showed less variation in activity patterns among months and sex-age classes, while adult males were observed more frequently during the autumn copulation period, suggesting that reproductive motivation affects their activity patterns.

Ten-year outcome of unilateral leg replantation after bilateral lower leg amputation following traumatic injury: A case report.

The absolute indications for lower limb replantation are not unequivocally established; rather, this procedure is still challenging and controversial. We report a case of a young male who underwent bilateral leg amputation, followed by unilateral replantation. The patient demonstrated good 10-year outcomes. A 23-year-old man had both of his lower legs crushed by heavy machinery that fell from the back of a truck, leading to amputation of bilateral lower limbs. Although bilateral amputation was recommended due to severe contusion on both sides, the patient and his family strongly requested replantation of the right leg, and surgery was started approximately 3 h after the injury. In addition to the emergency replantation, six surgeries were performed thereafter. Five months after the first surgery, the patient was fitted with a left lower leg prosthesis and started gait training. He was discharged 8 months later. Ten years after the replantation, the patient is satisfied with his leg. He has a normal gait with a prosthesis and has integrated into society with no functional deficit. There are still no clear standards for replantation after leg amputation, and individual decisions must be made based on the severity of the injury and the patient's general condition. In this case, we respected the patient's strong will to preserve the right leg and decided on the treatment plan. As a result, the patient was highly satisfied, and the spared right leg facilitated the patient's physical and mental recovery.

Pre-regulation of the planar chirality of pillar[5]arenes for preparing discrete chiral nanotubes.

Regulating the chirality of macrocyclic host molecules and supramolecular assemblies is crucial because chirality often plays a role in governing the properties of these systems. Herein, we describe pillar[5]arene-based chiral nanotube formation via pre-regulation of the building blocks' chirality, which is different from frequently used post-regulation strategies. The planar chirality of rim-differentiated pillar[5]arenes is initially regulated by chiral awakening and further induction/inversion through stepwise achiral external stimuli. The pre-regulated chiral information is well stored in discrete nanotubes by interacting with a per-alkylamino-substituted pillar[5]arene. Such pre-regulation is more efficient than post-regulating the chirality of nanotubes.

Cryopreservation of Biologically Functional Submandibular Gland Rudiments from Fetal Mice.

BACKGROUND/AIM: Cryopreservation of cell lines has been widely used in the laboratory; however, cryopreservation of organs is still considered to be difficult. The submandibular gland (SMG) of fetal mice is one of the best-characterized organs. We investigated the conditions for cryopreserving SMG rudiments. MATERIALS AND METHODS: Embryonic day 13 SMG rudiments were cryopreserved with or without a cryoprotectant. They were thawed and incubated in DMEM/F12 medium. Moreover, the influence of EGF stimulation on the signaling cascade after frozen-thawing the rudiments was analyzed by Western blotting. RESULTS: When SMG rudiments were cryopreserved without a cryoprotectant, all cells in the rudiments died. However, the SMG rudiments that had been preserved in a cryoprotectant showed branching morphogenesis. Additionally, the responsiveness of signaling cascades to EGF did not differ between frozen with a cryoprotectant and non-frozen rudiments. CONCLUSION: Cryopreservation might be a useful technology for preserving tissues from small organs, such as fetal SMG rudiments.

Alteration of gene expression in mice after glaucoma filtration surgery.

To clarify the early alterations of gene expression using a mouse model of glaucoma filtration surgery, we carried out microarray expression analysis. Using BALB/c mice, a filtration surgery model was made by incision of the limbal conjunctiva, followed by the insertion of a 33G needle tip into the anterior chamber, and 11-0 nylon sutures. Subgroups of mice were treated intraoperatively with 0.4 mg/ml mitomycin-C (MMC). At day 3 after surgery the bleb was maintained. The bleb region tissue was sampled 3 days after the filtration surgery, and gene expression analysis was carried out using a mouse Agilent 8 × 60 K array. We found 755 hyperexpressed transcripts in the bleb region compared to control conjunctiva. The hyperexpressed transcripts included epithelial cell metaplasia-related (Il1b, Krt16, Sprr1b), inflammation-related (Ccl2, Il6) and wound healing-related (Lox, Timp1) genes. We also found downregulation of a goblet cell marker gene (Gp2) in the bleb conjunctiva. MMC treatment suppressed elastin (Eln) gene expression and enhanced keratinization-related gene expression (Krt1, Lor) in the bleb region. Our results suggest the importance of epithelial wound healing after filtration surgery, and this filtration surgery model will be a useful tool for further pathophysiological analysis.

Sequential Chiral Induction and Regulator-Assisted Chiral Memory of Pillar[5]arenes.

Chirality transfer is widely observed in life processes, and many artificial chiral transfer systems have been developed. In these systems, chiral information is transferred from chiral inducers to chiral acceptors by a direct chiral induction process and a direct chiral memorization process. We have developed ternary nondirect chiral transfer systems based on pillar[5]arenes, in which a third factor was introduced as a regulator. The planar chirality of an acceptor was induced and memorized by a chiral inducer with precise control by a regulator. In the chiral induction period, the feed sequence of the chiral inducer and regulator affected the chiral induction behavior of the chiral acceptor. The chiral memory ability of the acceptor was substantially improved by the combined action of the chiral inducer and regulator.

(Hexafluoroacetylacetonato)copper(I)-cycloalkyne complexes as protected cycloalkynes.

A protection method for cycloalkynes by the formation of (hexafluoroacetylacetonato)copper(i)-cycloalkyne complexes is disclosed. Various complexes having functional groups were efficiently prepared, which are easily purified by silica-gel column chromatography. Selective click reactions were realized through the complexation of cycloalkynes with copper.

Selective strain-promoted azide-alkyne cycloadditions through transient protection of bicyclo[6.1.0]nonynes with silver or gold.

Complexation of bicyclo[6.1.0]nonynes with a cationic silver or gold salt results in protection from a click reaction with azides. The cycloalkyne protection using the silver or gold salt enables selective strain-promoted azide-alkyne cycloadditions of diynes keeping the bicyclo[6.1.0]nonyne moiety unreacted.

Design and synthesis of small molecule-conjugated photoaffinity nanoprobes for a streamlined analysis of binding proteins.

We designed and synthesized a set of photoaffinity nanoprobes, which multivalently display a small molecule ligand and a photoreactive group on gold nanoparticles. Due to the typically hydrophobic nature of these two functional groups, a hydrophilic spacer was additionally introduced to co-functionalize the nanoprobes to maintain their dispersibility in aqueous buffer solutions. Photoaffinity labeling studies using the nanoprobes composed of different ratios of three functional groups showed that including high density of the spacer group attenuates crosslinking efficiency. Comparative analysis of the reactivity among three major photoreactive groups suggested that unlike in the context of conventional photoaffinity probes, arylazide group enables the most selective crosslinking of a model small molecule binding protein.

Prevalence of High-Intensity Zones in the Lumbar Spine According to Age and Their Correlation with Other Degenerative Findings on Magnetic Resonance Imaging.

INTRODUCTION: A high-intensity zone (HIZ) in an intervertebral disc of the lumbar spine is a high-intensity signal located in the posterior annulus fibrosus on T2-weighted magnetic resonance imaging (MRI). There is limited information on the prevalence of HIZ in the lumbar spine according to age. The aim of this cross-sectional study was to investigate the prevalence of HIZ in the lumbar spine by age and the correlation between HIZ and other degenerative findings, such as disc degeneration, disc bulging and herniation, and changes in adjacent vertebral endplates on lumbar MRI. METHODS: We retrospectively reviewed MRI studies of 305 patients (1525 discs) with low back pain, leg pain, or numbness. The prevalence of HIZ was calculated in 5 age groups (<20, 20-39, 40-59, 60-79, 80-91 years). RESULTS: The number of patients in the 5 age groups was 19, 38, 69, 145, and 36, respectively. The prevalence of HIZ in the 5 age groups was 11.8%, 47.3%, 52.2%, 42.8%, and 50.0%, respectively. Disc degeneration was observed in 58.1% and 39.2% of discs with and without HIZ, respectively; disc bulging and herniation was observed in 63.9% and 41.1% and intensity changes at adjacent end plates in 11.6% and 10.0%, respectively. CONCLUSIONS: Prevalence of HIZ from the third decade of life onward was around 50%, with no significant change in prevalence beyond the age of 20 years. HIZ was correlated with disc degeneration, disc bulging, and disc herniation in patients with LBP, leg pain, or numbness.

Prognostic value of dynamic MRI positive enhancement integral color mapping in osteosynthesis of undisplaced femoral neck fractures.

AIMS: The common treatment for an undisplaced femoral neck fracture is osteosynthesis. Two major complications of osteosynthesis are non-union and late collapse of the femoral head. We speculated that femoral head perfusion is one of the most important factors that affect the outcome of osteosynthesis after femoral neck fracture. We have preoperatively estimated femoral head perfusion by dynamic MRI positive enhancement integral color mapping (PEICM). The purpose of this study was to evaluate the outcomes of undisplaced femoral neck fractures based on PEICM. PATIENTS AND METHODS: Sixty-eight patients participated in this prospective study. All patients underwent PEICM in a 1.5-Tesla MRI machine using coronal fast spoiled gradient echo imaging sequences with gadopentetate dimeglumine as the contrast agent. Femoral head perfusion was displayed via color mapping using PEICM. Three types were distinguished. For type A, the color was identical to unaffected side indicated normal perfusion. For type B, the color was darker than unaffected side indicated decreased perfusion. For type C, the color was black indicated complete absence of perfusion. All patients underwent osteosynthesis with three cannulated screws. The rates of non-union and late collapse for each type were calculated. RESULTS: Sixteen patients were classified as Type A, 43 as Type B, and 6 as Type C. The non-union rates were 0% for Type A, 6.7% for Type B, and 50.0% for Type C. The late collapse rates were 0% for Type A, 4.4% for Type B, and 0% for Type C. CONCLUSION: PEICM precisely detected femoral head perfusion. Primary prosthetic replacement should be considered for older patients with Type C to minimize the chances of revision surgery, even in undisplaced femoral neck fractures.

Radiographic changes of cervical destructive spondyloarthropathy in long-term hemodialysis patients: A 9-year longitudinal observational study.

Analyses of radiographic changes and clinical symptom of destructive spondyloarthropathy (DSA) on consecutive 42 patients managed with long-term hemodialysis were performed to elucidate radiographic changes of DSA and the factors that influence to the destructive changes. Patients underwent plain radiographs of the cervical spine with 9 years interval. Grading of radiological feature from lateral view was classified into grade 0 to grade 3. Clinical symptom was evaluated using modified Japanese Orthopaedic Association scoring system for cervical myelopathy (mJOA score). Destructive changes were observed in 3 patients at the first examination, and those were observed in 15 patients 9 years after the first examination. There is no statistically significant difference between the duration of hemodialysis and the grade. The mean age at the onset of hemodialysis, however, was significantly higher in patients of grade 2 and 3 than those of grade 1. Older patients with long-term hemodialysis had destructive changes. Destructive changes commonly observed in lower cervical spine. The average numbers of the involved disc level were 1.6 in grade 2 and 1.0 in grade 3. Clinical symptoms were varied in each grade and there was no statistically significant difference in total mJOA score among these grades. J. Med. Invest. 64: 68-73, February, 2017.

Incidence and clinical features of sacral insufficiency fracture in the emergency department.

INTRODUCTION: A sacral insufficiency fracture (SIF) often manifests as low back pain or sciatica in the absence of any antecedent trauma. These fractures may be missed because of lack of appropriate imaging. The purpose of this study was to clarify the incidence and clinical features of SIF as well as the characteristic findings on magnetic resonance imaging (MRI) of the lumbar spine. MATERIALS AND METHODS: The study participants comprised 250 patients (132 male, 118 female; mean age 58.6years) with pelvic trauma. SIF was identified on computed tomography or MRI. The incidence, initial symptoms, and time delay between the first visit and an accurate diagnosis of SIF were recorded. RESULTS: We detected 11 cases of SIF. Initial symptoms of SIF were low back pain (36.4%), gluteal pain (63.6%), and coxalgia (18.2%). Two patients complained of both low back pain and gluteal pain. The mean delay between the first visit and an accurate diagnosis of SIF was 23.9days. This time interval was significantly longer than in patients with other types of pelvic fracture. Four patients underwent MRI targeting the lumbar spine to investigate their symptoms. In all 4 patients, the signal intensity on T1-weighted and fat-suppressed images of the second sacral segment was low and high, respectively. CONCLUSION: This study demonstrates that accurate diagnosis of SIF may be delayed because of difficulties in detecting this type of fracture on plain X-ray and the non-specific nature of the presenting complaints. Emergency physicians should keep SIF in mind when investigating patients who complain of low back pain or gluteal pain. Findings at the second sacral segment on MRI targeting the lumbar spine may aid early diagnosis of this type of pelvic fracture.

Complete Fracture-Dislocation of the Thoracolumbar Spine with No Critical Neurological Deficit: A Case Report.

Fractures at the thoracolumbar junction are the most common spinal column fractures. Among type C fractures in the Arbeitsgemeinschaft für Osteosynthesefragen Spine Classification, cases with complete fracture-dislocations of the spinal column often result in a critical neurological deficit despite surgical treatment. We present a case of an 18-year-old man who had a complete fracture-dislocation of the T12 vertebral body and multiple injuries following high-energy trauma but no critical neurological deficits. Because of active bleeding in the left thoracic cavity, the patient underwent open reduction of the T12 vertebral body and anterior spinal fusion of the T11-L1 vertebral bodies via an anterior approach between the T9 and T10 ribs within 24 h of the accident. Four months postoperatively, the patient could ambulate independently, with a slight disturbance of light touch. At 6 months postoperatively, plain computed tomography scans showed bony union of the T12 vertebral body. We postulated two reasons for the absence of critical neurological dysfunction: (1) spontaneous spinal canal sparing because of the fracture of the right superior articular process in the L1 vertebral body and (2) fracture morphology, that is, a rotational fracture with mild to moderately strong shearing stress to the dura mater.

Injury to the Obturator Artery During Periacetabular Osteotomy: A Case Report.

CASE: We describe a middle-aged woman who had undergone periacetabular osteotomy (PAO) on both hips successively within a two-year period. Although the first PAO was successfully performed on the right hip without any complications, injury to the obturator artery occurred during the subsequent PAO on the left hip. In this case, the obturator artery seemed to be injured at the medial base of the iliopectineal eminence during osteotomy. CONCLUSION: PAO has been employed as a reconstructive surgery for acetabular dysplasia. However, given the proximity of intrapelvic vascular structures, there is a risk of iatrogenic vascular injury.

Cyclooxygenase-2 inhibitor inhibits the fracture healing.

We investigated the effects of cyclooxigenase-2 (cox-2) on fracture healing. After closed non-displaced fractures were created at the middle of both femoral shafts in 12-week-old Wister rats, a cox-2 specific inhibitor, etodolac (20 mg/day; intra-peritoneal) was administered every day for three weeks (E group). Bone union and callus formation were evaluated by weekly radiographs. Three weeks after surgery, the mechanical strength of the fractured femur was evaluated by a three-point-bending test. These results were compared with those of a vehicle control group (V group). The fracture healing score on radiographs in the E group three weeks after the surgery was 3.3 +/- 0.9, and in the V group it was 5.8 +/- 1.5, indicating that fracture healing was significantly poorer in the E than the V group (p < 0.05). From the three point bending test, the ultimate strength and stiffness of etodolac-treated fractured femurs were shown to be significantly lower than those in vehicle control group (p < 0.05). Mechanically, femurs of etodolac treated rats were weaker than those of control rats. Thus, it was concluded that etodolac, a cox-2 specific inhibitor, inhibited fracture healing.