Effective cultivation conditions and safety evaluation of filamentous cyanobacteria producing phycocyanins with antiglycation activities.

We investigated suitable culture conditions for the production of the blue pigment phycocyanin (PC) from the unique filamentous cyanobacteria Pseudanabaena sp. ABRG5-3 and Limnothrix sp. SK1-2-1. White, green, or red LED irradiation at 30 µmol photons/m2/s was effective for phycocyanin production when compared with Arthrospira platensis (Spirulina) sp. NIES-39, which is generally grown under high light irradiation. To investigate the safety of the cyanobacteria, ABRG5-3 cells were subjected to Ames (reverse mutation) tests and single oral-dose rat studies, which revealed non-mutagenic and non-toxic properties. When three purified phycocyanins (abPC, skPC, and spPC) were subjected to agarose gel electrophoresis, they showed different mobility, indicating that each phycocyanin has unique properties. abPC exhibited strong antiglycation activities as novel function.

Control of Relative Positions of Electron-Donor and Electron-Acceptor Molecules in Charge-Transfer Complexes for Luminescent Property Modulation.

Charge-transfer complexes can exhibit various physical properties that depend on the relative positions of electron-donor and electron-acceptor molecules. Several studies have investigated the relationship between the relative positions of electron-donor and electron-acceptor molecules and their luminescence properties. However, elucidating the correlation between the relative positions and detailed luminescence processes without changing the molecular structures has not been explored. Herein, we report control of the relative position based on charge-assisted hydrogen bonds between sulfo and amino groups and on alkylamines' steric factors, and report concomitant modulation of the luminescent properties. Six charge-transfer complexes were prepared from anthracene-2,6-disulfonic acid and 1,2,4,5-tetracyanobenzene as electron-donor and electron-acceptor molecules, and various alkylamines. Different alkylamines' steric factors drastically and precisely changed the relative positions of the electron-donor and electron-acceptor molecules without changing their molecular structures. Consequently, the six crystals exhibited maximum emission wavelengths from 543 to 624 nm and different luminescence processes.

Physical and psychological recovery after vaginal childbirth with and without epidural analgesia: A prospective cohort study.

BACKGROUND: Enhanced recovery is the gold standard in modern perioperative management, including that for cesarean deliveries. However, qualitative and quantitative data on the physical and psychological recovery of women after vaginal childbirth are limited. Whether neuraxial labor analgesia influences postpartum recovery is unknown. METHODS: Primiparous women anticipating a vaginal childbirth between January 2020 and May 2021 were enrolled. Women with major comorbidities or postpartum complications and those who underwent a cesarean delivery were excluded. Daily step count was measured using a wrist-worn activity tracker (FitbitTM Inspire HR) for 120 hours after vaginal childbirth. Subjective fatigue levels and health-related quality of life were assessed using the Multidimensional Fatigue Inventory (MFI) and EuroQol 5 Dimension 5 Level (EQ-5D-5L), respectively, at the 3rd trimester antenatal visit, on postpartum day 1 and 3, and at the one-month postpartum visit. Rest and dynamic pain scores and the location of pain were documented by participants during postpartum hospitalization. RESULTS: Among 300 women who were enrolled antenatally, 95 and 116 had a vaginal delivery without (NCB group) and with (EPL group) epidural analgesia, respectively. The median number of steps per 24 hours increased daily in both groups, and no significant difference was detected between the groups. Postpartum pain was mild overall, with median rest and dynamic pain scores being less than 4 and similar between the groups. MFI and EQ-5D-5L scores were the worst on postpartum day 1 in both groups and gradually improved to antepartum level by the one-month postpartum visit. Higher MFI score on postpartum day 1, but not the use of epidural analgesia, was associated with lower odds of achieving adequate postpartum ambulation (defined as >3500 steps between 48 and 72 hours postpartum). CONCLUSION: The use of epidural analgesia was not associated with worse recovery outcomes during postpartum hospitalization. TRIAL REGISTRATION: UMIN-CTR, #UMIN000039343, registered on January 31, 2020.

Influence of periostin on the development of fibrocartilage layers of anterior cruciate ligament insertion.

BACKGROUND: Periostin (Postn) is thought to play a role in the formation of anterior cruciate ligament (ACL) insertion. However, the influence of Postn on the development of ACL insertion requires further understanding. This study aimed to clarify the influence of Postn on the development of fibrocartilage layers of ACL insertion. HYPOTHESIS: We hypothesized that Postn would influence the development of fibrocartilage layers of ACL insertion. MATERIALS AND METHODS: C57BL/6N wild-type (Postn+/+; n=54) and Postn knockout (Postn-/-; n=54) mice were used in this study. Six animals were euthanized at 1 d and 1, 2, 3, 4, 6, 8, 10, and 12 weeks of age in each group. The chondrocyte number, proliferation, apoptosis, safranin O-stained glycosaminoglycan (GAG) area, type II collagen staining area, tidemark length, and insertion width were evaluated. RESULTS: Chondrocyte proliferation was high up to 2 weeks in Postn+/+, while low at age 1 d; it was, especially lower in Postn-/- than in Postn+/+ at age 1 d and 1 week. Chondrocyte apoptosis was high up to age 8 weeks in Postn+/+ and at 6 weeks in Postn-/-; it was especially higher in Postn-/- than in Postn+/+ at age 1 week. The GAG stained area was thickest for age 1 d to 4 weeks in Postn+/+ and for age 2 to 6 weeks in Postn-/-. The type II collagen staining area in Postn+/+ was thicker than that in Postn-/- at age 6 and 8 weeks. The tidemark length in Postn+/+ was longer than that in Postn-/- from age 8 to 12 weeks. The insertion width in Postn+/+ was longer than that in Postn-/- from age 1 to 3 weeks. DISCUSSION: Postn decreased cell proliferation in the early postnatal phase and influenced the development of the fibrocartilage layer extracellular matrix of ACL insertion in mice. Postn may contribute to the development of methods for regeneration of the ACL insertion. LEVEL OF EVIDENCE: V; controlled laboratory study.

Periostin Contributes to Fibrocartilage Layer Growth of the Patella Tendon Tibial Insertion in Mice.

Background and Objectives: The influence of periostin on the growth of the patella tendon (PT) tibial insertion is unknown. The research described here aimed to reveal the contribution of periostin to the growth of fibrocartilage layers of the PT tibial insertion using periostin knockout mice. Materials and Methods: In both the wild-type (WD; C57BL/6N, periostin +/+; n = 54) and periostin knockout (KO; periostin -/-; n = 54) groups, six mice were euthanized on day 1 and at 1, 2, 3, 4, 6, 8, 10, and 12 weeks of age. Chondrocyte proliferation and apoptosis, number of chondrocytes, safranin O-stained glycosaminoglycan (GAG) area, staining area of type II collagen, and length of the tidemark were investigated. Results: Chondrocyte proliferation and apoptosis in KO were lower than those in WD on day 1 and at 1, 4, and 8 weeks and on day 1 and at 4, 6, and 12 weeks, respectively. Although the number of chondrocytes in both groups gradually decreased, it was lower in KO than in WD on day 1 and at 8 and 12 weeks. In the extracellular matrix, the GAG-stained area in KO was smaller than that in WD on day 1 and at 1, 4, 8, 10, and 12 weeks. The staining area of type II collagen in KO was smaller than that in WD at 8 weeks. The length of the tidemark in KO was shorter than that in WD at 4 and 6 weeks. Conclusion: Loss of periostin led to decreased chondrocyte proliferation, chondrocyte apoptosis, and the number of chondrocytes in the growth process of the PT tibial insertion. Moreover, periostin decreased and delayed GAG and type II collagen production and delayed tidemark formation in the growth process of the PT tibial insertion. Periostin can, therefore, contribute to the growth of fibrocartilage layers in the PT tibial insertion. Periostin deficiency may result in incomplete growth of the PT tibial insertion.

Slowly progressive interstitial lung disease preceding typical dermatomyositis symptoms in anti-melanoma differentiation-associated gene 5 antibody-positive clinically amyopathic dermatomyositis.

A 73-year-old woman who visited our hospital complaining of dry cough for three months was refractory to antimicrobial therapy. Chest computed tomography revealed subpleural consolidation. Specimens obtained from surgical lung biopsy revealed subpleural perilobular airspace organization and fibrosis. After the biopsy, mechanic's hand and Gottron's papules appeared, and anti-melanoma differentiation-associated gene 5 (MDA5) antibody was found to be positive. Subsequently, anti-MDA5 antibody measured in cryopreserved serum from her first admission proved to be positive. It is difficult to suspect the presence of anti-MDA-5 antibody in patients with interstitial lung disease without typical dermatomyositis symptoms or slow disease progression.

Development of fibrocartilage layers in Achilles tendon enthesis in rabbits.

Objective: The details regarding the development of fibrocartilage layers in Achilles tendon (AT) enthesis are unknown. Therefore, we evaluated the development of fibrocartilage layers in AT enthesis using a rabbit model. Materials and Methods: Forty-eight male Japanese white rabbits were used in this study. Six of them were euthanized at different stages (day 1, and 1, 2, 4, 6, 8, 12, and 24 weeks of age). The proliferation, apoptosis, Sox9-positivity rates, and chondrocyte number were evaluated. Additionally, safranin O-stained glycosaminoglycan (GAG) areas, width of AT enthesis, and calcaneus length were assessed. All parameters were compared to those at 24 weeks of age. Results: The level of chondrocyte apoptosis was high from 1 to 8 weeks of age, and high expression level of Sox9 was maintained from day 1 to 6 weeks of age, which decreased gradually. Safranin O-stained GAG areas increased up to 12 weeks, calcaneus length increased up to 6 weeks, and the width of AT enthesis increased up to 1 week of age. Conclusion: The changes in chondrocyte and extracellular matrix were completed by 8 and 12 weeks of age, respectively. The development of fibrocartilage layers in AT enthesis was completed by 12 weeks of age. Our results contribute to the administration of appropriate treatments based on age and aid in the development of novel methods for regenerating AT enthesis.

Successful measures to prevent the spread of bovine papular stomatitis in a dairy farm.

Nasal papules and oral ulcers were observed in calves that were group-housed at a dairy farm. The calves were diagnosed with bovine papular stomatitis (BPS) due to parapoxvirus (PPV) infection based on virologic examinations using polymerase chain reaction to detect PPV. To prevent the spread of BPS, we isolated the affected calves, made procedural changes so that the affected herd was managed after the healthy herd, disinfected the bedding with slaked lime, disinfected the stalls and fences with invert soap, and changed the animals' feed to soft grass which does not damage the oral cavity. As a result, we succeeded in control the infection quickly.

Differences in the Development of Fibrocartilage Layers in the Quadriceps Tendon and Patellar Tendon Insertions in Rabbits: A Quantitative Study.

BACKGROUND: Differences in the development of fibrocartilage layers in quadriceps tendon (QT) and patellar tendon (PT) insertion sites are unclear. Because the mechanical environments for the QT and PT are different, the development of the QT and PT insertions may differ. PURPOSE: To investigate differences in the development of fibrocartilage layers in the QT and PT insertion sites in rabbits through use of quantitative morphometric evaluations. STUDY DESIGN: Descriptive laboratory study. METHODS: This study included 54 male Japanese White rabbits. Animals were euthanized at ages 1 day and 1, 2, 3, 4, 6, 8, 12, and 24 weeks (n = 6 for each age). Chondrocyte number, proliferation, apoptosis, sex-determining region Y box 9 (Sox9)-positive rates, safranin O-stained glycosaminoglycan (GAG) areas, tidemark length, insertion width, and patellar length were evaluated and compared with the same parameters at age 24 weeks and between QT and PT insertion sites. RESULTS: Chondrocyte proliferation was low up to age 2 weeks for QT insertion and low up to 1 week for PT insertion. Chondrocyte apoptosis was high at 1 day and Sox9 expression was low up to 1 week for PT insertion. Sox9 expression was higher in QT than in PT insertion at age 12 weeks. The high chondrocyte count continued to age 1 day in PT insertion and up to 6 weeks in QT insertion. The chondrocyte number was higher in QT than in PT insertion at age 2 weeks. The period of thicker GAG lasted from 2 to 8 weeks in PT insertion and from 1 to 12 weeks in QT insertion. GAG thickness in QT insertion was higher than in PT insertion at age 4 and 12 weeks. CONCLUSION: Development of fibrocartilage layers in QT and PT insertion sites was completed at age 24 weeks in rabbits. However, the period of high chondrocyte count and period of thicker GAG were longer in QT than in PT insertion up to 12 weeks. CLINICAL RELEVANCE: Development of fibrocartilage layers in QT and PT insertions differed in rabbits. Our results may contribute to the development of appropriate treatments based on age and the development of methods for regeneration of the insertion.

Novel predictive factors for patient discomfort and severe cough during bronchoscopy: A prospective questionnaire analysis.

During bronchoscopy, discomfort is mainly caused by an unavoidable cough; however, there are no reports of any predictive factors for strong cough during bronchoscopy identified before the procedure. To clarify the factors underlying the discomfort status and predictive factors for strong cough during bronchoscopy, we prospectively evaluated patients who underwent bronchoscopy at Kyorin University Hospital between March 2018 and July 2019. Before and after bronchoscopy, the enrolled patients answered a questionnaire regarding the procedure. At the same time, bronchoscopists evaluated cough severity using a four-grade cough scale. We evaluated patient characteristics and predictive factors associated with bronchoscopy from the perspective of discomfort and strong cough. A total of 172 patients were ultimately enrolled in this study. On multivariate logistic regression analysis, comparison of the subjective data between the discomfort and comfort groups revealed that factors that were more common in the former group were younger age (OR = 0.96, p = 0.002), less experienced bronchoscopist (OR = 2.08, p = 0.047), and elevation of cough score per 1 point (OR = 1.69, p < 0.001). Furthermore, the predictive factors for strong cough prior to performing bronchoscopy were female sex (OR = 2.57, p = 0.009), EBUS-TBNA (OR = 2.95, p = 0.004), and prolonged examination time of more than 36 min (OR = 2.32, p = 0.022). Regarding patients' discomfort, younger age, less experienced bronchoscopist, and the elevation of cough score per 1 point were important factors for discomfort in bronchoscopy. On the other hand, female sex, EBUS-TBNA, and prolonged examination time were crucial factors for strong cough.

Development of fibrocartilage layers in the anterior cruciate ligament insertion in rabbits.

BACKGROUND: A detailed evaluation focusing on the fibrocartilage layers in the anterior cruciate ligament (ACL) insertion is necessary to consider regeneration of the insertion. This study examined the development of the fibrocartilage layers in the ACL tibial insertion in rabbits by quantitative morphometric evaluations based on histological and immunohistochemical analyses. METHODS: Male Japanese white rabbits were used because of their history of use for histomorphometric analyses of the ACL insertion and to eliminate the influence of female hormones on the ACL. Six animals were euthanized at each age (1 day and 1, 2, 4, 6, 8, 12, and 24 weeks); in total, 48 animals were used. Proliferation rate, apoptosis rate, Sox9-positive rate, and chondrocyte number were evaluated. Safranin O-stained glycosaminoglycan (GAG) areas, tidemark length, ACL insertion width, and ACL length were also evaluated. All parameters were compared with those at age 24 weeks of age. RESULTS: High levels of chondrocyte proliferation and Sox9 expression continued until 4 and 8 weeks of age, respectively, and then gradually decreased. Chondrocyte apoptosis increased up to 8 weeks. The chondrocyte number, ACL insertion width, ACL length, safranin O-stained GAG areas, and tidemark length gradually increased up to 12 weeks. CONCLUSION: Chondrocytes that displayed chondrocyte proliferation and Sox9 expression increased until 12 weeks of age, in accordance with development of the ACL length and its insertion width. The GAG production and tidemark length also increased until 12 weeks of age. The development of fibrocartilage layers in the ACL insertion was complete at 12 weeks of age.

Extension of knee immobilization delays recovery of histological damages in the anterior cruciate ligament insertion and articular cartilage in rabbits.

[Purpose] To investigate the influence of knee immobilization period on recovery of histological damages in the anterior cruciate ligament (ACL) insertion and articular cartilage in rabbits. This knowledge is important for determining the appropriate rehabilitation approach for patients with ligament injuries, fracture, disuse atrophy, and degenerative joint disease. [Materials and Methods] Forty-eight male Japanese white rabbits were divided equally into the remobilization and control groups. The remobilization group had the right knee surgically immobilized, and was divided equally into four subgroups according to the duration of immobilization (1, 2, 4 and 8 weeks). After the immobilization was removed, the rabbits moved freely for 8 weeks. The control group underwent sham operation and followed the same time course as the remobilization group. The chondrocyte apoptosis rate and chondrocyte proliferation rate in the ACL insertion and articular cartilage were analyzed after remobilization. [Results] In the ACL insertion, the remobilization group had a higher chondrocyte apoptosis rate than the control group after 8 weeks of immobilization, and a lower chondrocyte proliferation rate than the control group after 4 and 8 weeks of immobilization. In the articular cartilage, the remobilization group had a lower chondrocyte proliferation rate than the control group after 8 weeks of immobilization. After 8 weeks of remobilization, the ACL insertion and articular cartilage are not completely recovered after 4 and 8 weeks of immobilization, respectively. [Conclusion] Our results suggest that 8 weeks of remobilization will result in recovery of the ACL insertion after 2 weeks of knee immobilization, and recovery of the articular cartilage after 4 weeks of knee immobilization. If 8 weeks of immobilization occurs, a remobilization duration of more than 8 weeks may be necessary.

Differences in placement of calcium phosphate-hybridized tendon grafts within the femoral bone tunnel during ACL reconstruction do not influence tendon-to-bone healing.

BACKGROUND: Calcium phosphate (CaP)-hybridization of tendon grafts has been shown to improve tendon-to-bone healing. The purpose of this study was to clarify the influence of different tendon graft placement methods on tendon-to-bone healing using CaP-hybridized tendon grafts in anterior cruciate ligament (ACL) reconstructions in rabbits. METHODS: We compared two methods of tendon graft placement within the femoral bone tunnel: suspension of the tendon graft within the bone tunnel (suspension group) and implantation of the tendon graft coherent with the bone socket (coherence group). CaP-hybridized tendon grafts were used in both groups. Fifty-six male Japanese white rabbits were used for this study. The results of biomechanical tests (n = 9) and histological analyses (n = 5) were evaluated at 2 and 4 weeks after surgery. RESULTS: The ultimate failure load, stiffness, stress, soft tissue remaining in bone tunnel after biomechanical testing, and direct bonding area at tendon-bone interface did not differ significantly between the suspension and coherence groups at either 2 or 4 weeks after surgery (p > 0.05). In both groups, the ultimate failure load, stress, soft tissue remaining in the bone tunnel, and direct bonding area at interface at 4 weeks after surgery were significantly greater than those at 2 weeks after surgery (p < 0.05). CONCLUSIONS: Tendon-to-bone healing in both groups progressed until the endpoint of 4 weeks. There was no influence of the CaP-hybridized tendon graft placement method on tendon-to-bone healing at 4 weeks after ACL reconstruction in rabbits. Thus, the CaP-hybridized tendon grafts were unaffected by differences in their placement within the bone tunnel and became equally anchored to the bone tunnel during the early postoperative period. The tendon graft placement method may not influence tendon-to-bone healing in ACL reconstruction when CaP-hybridized tendon grafts are used.

Influence of Knee Immobilization on Chondrocyte Apoptosis and Histological Features of the Anterior Cruciate Ligament Insertion and Articular Cartilage in Rabbits.

This study examined the influence of immobilization on chondrocyte apoptosis and histological features of the anterior cruciate ligament (ACL) insertion and knee articular cartilage in rabbits. Forty-eight male Japanese white rabbits were assigned to an immobilization (n = 24) or sham (n = 24) group. Rabbits in the immobilization group underwent complete unilateral surgical knee immobilization and rabbits in the sham group underwent a sham surgery. The average thickness of the glycosaminoglycan (GAG) stained red area by safranin O staining, the chondrocyte apoptosis rate and the chondrocyte proliferation rate in the cartilage layer in the ACL insertion and the articular cartilage of the medial tibial condyle were measured at one, two, four and eight weeks in six animals from each group. In the ACL insertion, the chondrocyte apoptosis rate was higher in the immobilization group than in the sham group at two and eight weeks after surgery (p < 0.05). The chondrocyte proliferation rate gradually decreased from two weeks to eight weeks in the immobilization group. The GAG layer was thinner in the immobilization group than in the sham group at two, four and eight weeks after surgery (p < 0.05). In the articular cartilage, the chondrocyte apoptosis rate in the immobilization group was higher than in the sham group at four and eight weeks after surgery (p < 0.05). The GAG layer was significantly thinner in the immobilization group than that in the sham group at four and eight weeks after surgery (p < 0.05). Knee immobilization significantly increased chondrocyte apoptosis at two and eight weeks after surgery in the ACL insertion and at four and eight weeks after surgery in the articular cartilage of the medial tibial condyle, and decreased GAG layer thickness from two to eight weeks after surgery in the ACL insertion and from four to eight weeks after surgery in the articular cartilage.

Effect of Calcium Phosphate-Hybridized Tendon Graft in Anatomic Single-Bundle ACL Reconstruction in Goats.

BACKGROUND: We previously developed a novel technique using an alternate soaking process that improves tendon-bone healing by hybridizing the tendon graft with calcium phosphate (CaP). However, the effects of the CaP-hybridized tendon graft on anatomic single-bundle anterior cruciate ligament (ACL) reconstruction remain unclear. PURPOSE: To determine the effects of CaP-hybridized tendon grafts compared with untreated tendon grafts 6 months after anatomic single-bundle ACL reconstruction using a goat model. STUDY DESIGN: Controlled laboratory study. METHODS: Animals were divided into a CaP group (n = 5 goats) and a control group (n = 5 goats), and we analyzed (1) knee kinematics and in situ forces under applied anterior tibial loads of 50 N and internal tibial torque of 2.0 N·m in the grafts at full extension and at 60° and 90° of knee flexion, (2) the mean percentage of bone tunnel enlargement using computed tomography (CT), and (3) the histology of the tendon-bone interface. RESULTS: The in situ forces under applied anterior tibial loads of 50 N at 60° and 90° of knee flexion in the CaP group were greater than those in the control group (P < .05). The red safranin-O-stained area, indicating glycosaminoglycans in the cartilage layers at the joint aperture sites of the anterior femoral and posterior tibial bone tunnel, was greater in the CaP group than that in the control group (P < .05). The lengths of the nonbonding gap area between the anterior femoral and posterior tibial bone tunnels in the control group were greater than those in the CaP group (P < .05). No significant difference could be detected in the mean percentage of bone tunnel enlargement between the 2 groups. CONCLUSION: The CaP-hybridized tendon graft enhanced tendon-bone healing at the joint aperture site in both anterior femoral and posterior tibial tunnels 6 months after anatomic single-bundle ACL reconstruction in goats. The in situ forces under applied anterior tibial loads at greater flexion angles in the CaP group increased compared with controls. CLINICAL RELEVANCE: Anatomic single-bundle ACL reconstruction using CaP-hybridized tendon grafts may lead to better postoperative knee function.

Drug-susceptibility of isolates of Brachyspira hyodysenteriae isolated from colonic mucosal specimens of pigs collected from slaughter houses in Japan in 2009.

Twenty nine isolates identified as Brachyspira hyodysenteriae were most susceptible to carbadox and metronidazole, whereas they were resistant to macrolides. The isolates showed intermediate susceptibility to tiamulin, lincomycin, penicillin G, ampicillin, chloramphenicol, tetracycline, enrofloxacin and valnemulin, with MIC50 values ranging from 0.39 to 3.13.

Influence of mechanical unloading on histological changes of the patellar tendon insertion in rabbits.

BACKGROUND: The purpose of this study was to clarify the influence of mechanical unloading on histological changes of the patellar tendon (PT) insertion in rabbits. MATERIALS AND METHODS: The PT was completely released from stress by drawing the patella toward the tibial tubercle with a stainless steel wire installed between the patella and tibial tubercle (mechanical unloading group, n=28). The animals of the sham group underwent the same surgical procedure; however, the wire was not tightened (n=28). The average thickness of the Safranin O-stained glycosaminoglycan (GAG) area, chondrocyte apoptosis rate and chondrocyte proliferation rate of the cartilage layer at the insertion were measured at one, two, four, and six weeks. RESULTS: The chondrocyte apoptosis rate in the mechanical unloading group was significantly higher than that in the sham group at one and four weeks (p<0.05). The chondrocyte proliferation rate in the mechanical unloading group was significantly lower than that in the sham group at four and six weeks (p<0.05). The average thickness of the GAG-stained area in the mechanical unloading group was significantly lower than that in the sham group at six weeks (p<0.05). CONCLUSION: Mechanical unloading significantly affected the increase in the chondrocyte apoptosis rate, decrease in the chondrocyte proliferation rate, and decrease in the GAG layer thickness at the PT insertion for up to six weeks in rabbits. CLINICAL RELEVANCE: We suggest that more than 6 weeks of mechanical unloading should be avoided to prevent degeneration at the PT insertion.

Influence of gradual elongation to the patella tendon insertion in rabbits.

The purpose of this study was to examine the histological changes at the patella tendon (PT) insertion site under gradual elongation in rabbits. Gradual elongation of the PT was performed using external fixation for 4 weeks, with a lengthening speed of 0.5 mm/day (elongation group; n = 24). Rabbits in the sham group underwent the same surgical procedure without gradual elongation (sham group; n = 24). Eight animals were sacrificed 1, 2 and 4 weeks after surgery in each group, respectively. Average thicknesses of stained glycosaminoglycan (GAGs) areas by Safranin-O staining in the total cartilage layer and the uncalcified fibrocartilage layer in the elongation group were significantly higher than that in the sham group at 4 weeks (p < 0.05) and that in the intact PT group (n = 6, p < 0.05). In the elongation group, the peak in the average thicknesses of the stained GAGs areas in the total cartilage layer and the uncalcified fibrocartilage layer were observed at 4 weeks. Gradual elongation of PT insertion significantly affected the increase in the average thicknesses of the stained GAGs areas in the cartilage layer especially in the uncalcified fibrocartilage layer at 4 weeks in rabbits. Clinically, insertions of tendon and ligament can extend during gradual elongation using external fixation more than 4 weeks after the operation.

[A case of emergency resection of a giant gastrointestinal stromal tumor of the stomach associated with hemorrhagic shock].

We report a case of emergency resection of a giant gastrointestinal stromal tumor of the stomach associated with hemorrhagic shock. A 79-year-old woman was admitted to our hospital because of massive hematemesis. Laboratory analysis revealed a hemoglobin level of 6.5 g/dL. Abdominal computed tomography (CT) and upper gastrointestinal endoscopy revealed a submucosal tumor, 12 cm in diameter, in the fornix of the stomach. As a Dieulafoy-like lesion was present, we attempted coagulation hemostasis in the exposed blood vessels. Endoscopic hemostasis was not successful. The patient went into hemorrhagic shock. Emergency surgery was performed: total gastrectomy with distal pancreatosplenectomy. The resected specimen measured 10×12×7 cm and was hard. Immunohistologically, the tumor was positive for c-kit and CD34 and negative for alpha smooth muscle actin (αSMA), desmin, and S-100. Histological examination revealed that the patient had a high-risk gastrointestinal stromal tumor of the stomach with no nodal metastasis. The postoperative course was uneventful and the patient has remained alive without recurrence for 4 years.

Anterior cruciate ligament insertion after partial tear: histological changes and chondrocyte turnover.

PURPOSE: The purpose of this study was to clarify the effects of partial resection on the glycosaminoglycan (GAG) layer thicknesses and chondrocyte turnover (apoptosis and cell proliferation) between uncalcified fibrocartilage (UF) and calcified fibrocartilage (CF) layers in an anterior cruciate ligament (ACL) insertion. METHODS: Twenty male Japanese white rabbits were evaluated. The anteromedial bundle of the ACL substance was resected in the right knee. The posterolateral bundle was left intact. Five rabbits were evaluated at 1, 2, 4, and 8 weeks after surgery, respectively. RESULTS: The apoptosis rates in the UF and CF layers were significantly lower in the posterolateral area than those in the anteromedial area at 1 and 2 weeks, respectively. The cell proliferation rates in the UF and CF layers were significantly higher in the posterolateral area than those in the anteromedial area at 2 and 4 weeks, respectively. The GAG layer thicknesses in the UF and CF layers were higher in the posterolateral area than those in the anteromedial area at 1-8 and 2-8 weeks, respectively. The GAG layer thicknesses in the UF and CF layers in the posterolateral area peaked at 2 and 4 weeks, respectively. However, the thicknesses in the two layers in the posterolateral area gradually decreased until 8 weeks. CONCLUSION: The GAG layer thicknesses in the UF and CF layers in the remaining ligament area increased up to 4 weeks and gradually decreased until 8 weeks owing to an imbalance between chondrocyte apoptosis and proliferation. If the reactions in humans are similar to those observed in the rabbits, we consider that augmentation for ligament reconstruction and partial repair should be performed within at least 1 month after injury, before insertion degeneration occurs.