Anatomical evaluation of the superficial medial collateral ligament distal tibial attachment of the knee.

This study aimed to evaluate the superficial medial collateral ligament distal tibial attachment (sMCL-dTA) morphologically and morphometrically. Seventeen unpaired formalin-fixed cadaveric knees were used. The sMCL was divided into anterior and posterior sections in the paracoronal plane along the midline of the sMCL. The distance from the medial edge of the tibial plateau and the joint line to the proximal margin, center, and distal margin of the sMCL-dTA and the length of the sMCL-dTA were measured in the anterior section, respectively. The sMCL-dTA was histologically observed in the posterior section with hematoxylin and eosin and Masson's trichrome staining. The distance from the medial edge of the tibial plateauto the proximal margin, center, and distal margin of the sMCL-dTA were 38.1 ± 4.2, 49.7 ± 4.4, and 61.5 ± 5.1 mm, respectively. The perpendicular distance from the joint line to the proximal margin, center, and distal margin of the sMCL-dTA were 36.1 ± 4.0, 47.4 ± 4.2, and 59.1 ± 4.8 mm, respectively. The length of the sMCL-dTA was 23.6 ± 3.2 mm. Histologically, the sMCL-dTA was formed by two layers of collagen fibers: the unidirectional fibrous layer and the multidirectional fibrous layer. The respective thicknesses of the two layers both decreased distally. The anatomical location, the length, and the attachment morphology of sMCL-dTA have been clarified using human cadaveric knees. Anatomical data in the present study contribute to the quality of surgery associated with sMCL-dTA.

A new injection method for identifying the subpopliteal recess of the knee.

The posterolateral region of the knee has a complex and diverse anatomy. Hydrarthrosis of the knee can potentially communicate with other parts of the joint space. The joint fluid distribution reflects anatomical communications between synovial spaces. To observe the continuity between the knee joint cavity and the surrounding bursa, we devised a dissection method with a new injection agent, an eosin-containing congealed liquid that spreads uniformly over the entire space. The purpose of this study was to perform a detailed examination of the subpopliteal recess (SPR) where a bursa connects to the knee joint capsule. We also reported the advantages of this new injection agent compared with conventional materials (latex and epoxy resin). Twenty-two formalin-fixed cadavers (34 knees), two N-vinyl-pyrrolidone (NVP)-fixed cadavers (4 knees), and two cadavers (3 knees) fixed by Thiel's method were used. After filling the knee joint space and SPR with eosin congealed liquid, the specimens were dissected to investigate the morphology of the SPR. In addition, three different types of injection agents were assessed. The SPR extended distally along the popliteus tendon. The SPR length was 22.64 ± 11.38 mm from the upper end of the lateral tibial condyle to the lower end of the depression. The existence of a fabellofibular ligament made the SPR significantly longer, but abrasion of the femoral articular cartilage did not affect the SPR. Furthermore, the relationship between the popliteus muscle and the SPR was classified into three types (types 1-3). Types 2 and 3 in which the SPR extended to the proximal tibiofibular joint may cause instability of the knee joint. The eosin congealed liquid was highly useful in many aspects, such as fluidity and injection workability. The new dissection method with eosin congealed liquid provides insights into the anatomy of the posterior lateral knee, which are useful for radiological diagnoses and clinical treatments.

Infusion of Variable Chemical Structure to Tune Stacking among Metal-Organic Layers in 2D Nano MOF.

Thickness of two-dimensional (2D) metal-organic frameworks (MOFs) govern their intriguing functionalities. Primarily this thickness is controlled by the stacking between the metal-organic layers (MOL). It is observed that until now such modulating factors for stacking efficiency of MOL are not well studied. Here, we report a fundamental hypothesis to comprehend regulation of stacking efficiency among MOLs as a function of chemical structure of organic ligands (dicarboxylic acids and pillar linkers). This basically involves a series of isostructural three-dimensional (3D) MOFs which contain linkers of variable chemical nature that could be depillared to generate 2D stacked MOFs of different thickness. Depending on the linkers, we encountered the formation of single MOL to stacked multiple MOLs as evidenced from atomic force microscopic and other experimental analysis. The present study gives a concrete correlation between the stacking within 2D MOFs (from monolayer to multilayers), and their 3D counter parts, which may provide a thickness tuning pathway for 2D MOFs.

Infusion of Variable Chemical Structure to Tune Stacking among Metal-Organic Layers in 2D Nano MOFs.

Invited for the cover of this issue is the group of Biplab Manna at the University of Kumamoto. The image depicts various kinds of stacking between the metal-organic layers of MOFs. Read the full text of the article at 10.1002/chem.202201665.

Replication of fouling in vitro in hollow fiber dialyzers by albumin immobilization.

For designing and evaluating the dialyzer and investigating the optimal therapeutic conditions, in vitro studies bring us many useful findings. In hemodialysis, however, the membrane fouling due to protein molecules reduces solute removal performance. Therefore, we investigated a method for replicating the fouling in dialyzers in aqueous experiments. After the albumin solution was circulated in the test circuit with a dialyzer, a glutaraldehyde solution was pumped into the dialyzer to immobilize albumin on the hollow fiber membrane. Under various immobilization conditions, the permeability of creatinine and vitamin B12 was evaluated by dialysis experiments. The creatinine clearance after immobilization of albumin was decreased, suggesting pore plugging by our fouling replication method. The glutaraldehyde crosslinked albumin molecules that adhered them to the membrane firmly. Moreover, the degree of fouling may be controlled by changing the concentration of albumin solution and the volume of glutaraldehyde solution used for immobilization. Our fouling replication method was applied to three types of polyester polymer alloy (PEPA) dialyzers and one polysulfone (PSf) dialyzer. This method enables to evaluate the permeability of various dialyzers with fouling in vitro that will be of great help in collecting data for designing dialyzers.

Novel substitution technique in intermittent infusion hemodiafiltration (I-HDF) therapy using back filtration as substitution.

An infusion of dialysate into the blood compartment across the membrane using back filtration in dialysis therapy provides a stabilizing blood pressure and a membrane flushing during treatment. We devised a method to flush the membrane effectively and tried to find the optimum infusion patterns for intermittent infusion hemodiafiltration (I-HDF) from the aspect of solute removal by computing the pressure distribution in a diafilter. Bovine blood experiments were performed under following three modes: control HD in which no intentional filtration was involved, and two I-HDF in which back filtration was made either under counter current or under parallel flow. The inner surface of the hollow fiber before and after the experiment was observed using FE-SEM. According to the computation of the pressure distribution, a large amount of normal filtration occurs near the blood inlet in control HD. In addition, when the back filtration is performed under parallel flow, the amount of backfiltration near the blood inlet is 3.43 times higher than that in the case of counter current. Clearance (CL) of inulin remained at the highest level when the back filtration was performed under parallel flow. Near the blood inlet where the fouling was significantly formed, many macropores remained on the membrane when the backfiltration was performed under parallel flow. The degree of fouling showed a distribution along with the blood flow and the pressure distribution. Furthermore, the more effective recovery of CL can be expected by introducing the backfiltration under parallel flow to which fouling was significantly formed.

Recent progress of animal transplantation studies for treating articular cartilage damage using pluripotent stem cells.

Focal articular cartilage damage can eventually lead to the onset of osteoarthritis with degradation around healthy articular cartilage. Currently, there are no drugs available that effectively repair articular cartilage damage. Several surgical techniques exist and are expected to prevent progression to osteoarthritis, but they do not offer a long-term clinical solution. Recently, regenerative medicine approaches using human pluripotent stem cells (PSCs) have gained attention as new cell sources for therapeutic products. To translate PSCs to clinical application, appropriate cultures that produce large amounts of chondrocytes and hyaline cartilage are needed. So too are assays for the safety and efficacy of the cellular materials in preclinical studies including animal transplantation models. To confirm safety and efficacy, transplantation into the subcutaneous space and articular cartilage defects have been performed in animal models. All but one study we reviewed that transplanted PSC-derived cellular products into articular cartilage defects found safe and effective recovery. However, for most of those studies, the quality of the PSCs was not verified, and the evaluations were done with small animals over short observation periods. Large animals and longer observation times are preferred. We will discuss the recent progress and future direction of the animal transplantation studies for the treatment of focal articular cartilage damages using PSCs.

Electrophysiological insights into the relationship between calcium dynamics and cardiomyocyte beating function in chronic hemodialysis treatment.

For patients in which the Ca2+ concentration of dialysis fluid is lower than that in plasma, chronic hemodialysis treatment often leads to cardiac beating dysfunction. By applying these conditions to an electrophysiological mathematical model, we evaluated the impact of body fluid Ca2+ dynamics during treatment on cardiomyocyte beating and, moreover, explored measures that may prevent cardiomyocyte beating dysfunction. First, Ca2+ concentrations in both plasma and interstitial fluid were decreased with treatment time, which induced both a slight decline in beating rhythm on a sinoatrial nodal cell and a wane in contraction force on a ventricular cell. These simulated results were in agreement with clinical observations. Next, a relationship between the intracellular Ca2+ concentration and ion current dynamics of ion transporters were examined to elucidate the mechanism underlying cardiomyocyte beating dysfunction. The inward current of the Na/Ca exchanger (NCX) increased with a decrease in Ca2+ concentration in interstitial fluid and induced a reduction in intracellular Ca2+ concentration during treatment. Furthermore, the decline in intracellular Ca2+ concentration reduced the contraction force. These findings implied that ion transport through the NCX is a dominant factor that induces cardiomyocyte beating dysfunction during hemodialysis. Finally, the replenishment of Ca2+ or application of an NCX inhibitor during treatment suppressed the decrease in intracellular Ca2+ concentration and contributed to the stabilization of cardiomyocyte beating function. In summary, the clinical implementation of hepatically cleared NCX inhibitor may be a suitable approach to improving the quality of life for patients on chronic hemodialysis.

Statin treatment rescues FGFR3 skeletal dysplasia phenotypes.

Gain-of-function mutations in the fibroblast growth factor receptor 3 gene (FGFR3) result in skeletal dysplasias, such as thanatophoric dysplasia and achondroplasia (ACH). The lack of disease models using human cells has hampered the identification of a clinically effective treatment for these diseases. Here we show that statin treatment can rescue patient-specific induced pluripotent stem cell (iPSC) models and a mouse model of FGFR3 skeletal dysplasia. We converted fibroblasts from thanatophoric dysplasia type I (TD1) and ACH patients into iPSCs. The chondrogenic differentiation of TD1 iPSCs and ACH iPSCs resulted in the formation of degraded cartilage. We found that statins could correct the degraded cartilage in both chondrogenically differentiated TD1 and ACH iPSCs. Treatment of ACH model mice with statin led to a significant recovery of bone growth. These results suggest that statins could represent a medical treatment for infants and children with TD1 and ACH.

Changes in acetyl-CoA mediate Sik3-induced maturation of chondrocytes in endochondral bone formation.

The maturation of chondrocytes is strictly regulated for proper endochondral bone formation. Although recent studies have revealed that intracellular metabolic processes regulate the proliferation and differentiation of cells, little is known about how changes in metabolite levels regulate chondrocyte maturation. To identify the metabolites which regulate chondrocyte maturation, we performed a metabolome analysis on chondrocytes of Sik3 knockout mice, in which chondrocyte maturation is delayed. Among the metabolites, acetyl-CoA was decreased in this model. Immunohistochemical analysis of the Sik3 knockout chondrocytes indicated that the expression levels of phospho-pyruvate dehydrogenase (phospho-Pdh), an inactivated form of Pdh, which is an enzyme that converts pyruvate to acetyl-CoA, and of Pdh kinase 4 (Pdk4), which phosphorylates Pdh, were increased. Inhibition of Pdh by treatment with CPI613 delayed chondrocyte maturation in metatarsal primordial cartilage in organ culture. These results collectively suggest that decreasing the acetyl-CoA level is a cause and not result of the delayed chondrocyte maturation. Sik3 appears to increase the acetyl-CoA level by decreasing the expression level of Pdk4. Blocking ATP synthesis in the TCA cycle by treatment with rotenone also delayed chondrocyte maturation in metatarsal primordial cartilage in organ culture, suggesting the possibility that depriving acetyl-CoA as a substrate for the TCA cycle is responsible for the delayed maturation. Our finding of acetyl-CoA as a regulator of chondrocyte maturation could contribute to understanding the regulatory mechanisms controlling endochondral bone formation by metabolites.

Modeling type II collagenopathy skeletal dysplasia by directed conversion and induced pluripotent stem cells.

Type II collagen is a major component of cartilage. Heterozygous mutations in the type II collagen gene (COL2A1) result in a group of skeletal dysplasias known as Type II collagenopathy (COL2pathy). The understanding of COL2pathy is limited by difficulties in obtaining live chondrocytes. In the present study, we converted COL2pathy patients' fibroblasts directly into induced chondrogenic (iChon) cells. The COL2pathy-iChon cells showed suppressed expression of COL2A1 and significant apoptosis. A distended endoplasmic reticulum (ER) was detected, thus suggesting the adaptation of gene expression and cell death caused by excess ER stress. Chondrogenic supplementation adversely affected the chondrogenesis due to forced elevation of COL2A1 expression, suggesting that the application of chondrogenic drugs would worsen the disease condition. The application of a chemical chaperone increased the secretion of type II collagen, and partially rescued COL2pathy-iChon cells from apoptosis, suggesting that molecular chaperons serve as therapeutic drug candidates. We next generated induced pluripotent stem cells from COL2pathy fibroblasts. Chondrogenically differentiated COL2pathy-iPS cells showed apoptosis and increased expression of ER stress-markers. Finally, we generated teratomas by transplanting COL2pathy iPS cells into immunodeficient mice. The cartilage in the teratomas showed accumulation of type II collagen within cells, a distended ER, and sparse matrix, recapitulating the patient's cartilage. These COL2pathy models will be useful for pathophysiological studies and drug screening.

Support for Dialysis Therapy in Vietnam, Cambodia, and Myanmar by Japanese Societies in the Field of Blood Purification.

With recent economic development in Southeast Asia, there have been improvements in medical services and healthcare provision. This has led to increased numbers of dialysis patients and increased numbers of dialysis facilities in the region. To assist economically developing countries in managing this change, support projects from Japan have been conducted in the region since around 2007. This article summarizes and discusses Japan's support activities, in which some of the authors were directly involved, in Vietnam, Cambodia, and Myanmar. Initial support was mainly organized by the non-governmental organization Ubiquitous Blood Purification International (NGO UBPI), and currently several organizations in the field of blood purification work together to offer ongoing support in the region. Many positive changes have resulted from these activities in Southeast Asia, but challenges remaining for the future are to establish an educational system for each dialysis specialty and develop dialysis techniques ensuring high treatment quality and safety.

Prevalence of elevated serum anti-N-methyl-D-aspartate receptor antibody titers in patients presenting exclusively with psychiatric symptoms: a comparative follow-up study.

BACKGROUND: Increasing numbers of patients with elevated anti-N-methyl-D-aspartate (NMDA) receptor antibody titers presenting exclusively with psychiatric symptoms have been reported. The aim of the present study was to clarify the prevalence of elevated serum anti-NMDA receptor antibody titers in patients with new-onset or acute exacerbations of psychiatric symptoms. In addition, the present study aimed to investigate the association between elevated anti-NMDA receptor titers and psychiatric symptoms. METHODS: The present collaborative study included 59 inpatients (23 male, 36 female) presenting with new-onset or exacerbations of schizophrenia-like symptoms at involved institutions from June 2012 to March 2014. Patient information was collected using questionnaires. Anti-NMDA receptor antibody titers were measured using NMDAR NR1 and NR2B co-transfected human embryonic kidney (HEK) 293 cells as an antigen (cell-based assay). Statistical analyses were performed for each questionnaire item. RESULTS: The mean age of participants was 42.0 ± 13.7 years. Six cases had elevated serum anti-NMDA antibody titers (10.2 %), four cases were first onset, and two cases with disease duration >10 years presented with third and fifth recurrences. No statistically significant difference in vital signs or major symptoms was observed between antibody-positive and antibody-negative groups. However, a trend toward an increased frequency of schizophrenia-like symptoms was observed in the antibody-positive group. CONCLUSION: Serum anti-NMDA receptor antibody titers may be associated with psychiatric conditions. However, an association with specific psychiatric symptoms was not observed in the present study. Further studies are required to validate the utility of serum anti-NMDA receptor antibody titer measurements at the time of symptom onset.

Derivation of iPSCs in stirred suspension bioreactors.

Induced pluripotent stem cells (iPSCs) are typically derived in adherent culture. Here we report fast and efficient derivation of mouse iPSCs in stirred suspension bioreactors, with and without the use of c-Myc. Suspension-reprogrammed cells expressed pluripotency markers, showed multilineage differentiation in vitro and in vivo, and contributed to the germline in chimeric mice. Suspension reprogramming has the potential to accelerate and standardize iPSC research.

Subpopulations of fibroblasts derived from human iPS cells.

Organ fibrosis causes collagen fiber overgrowth and impairs organ function. Cardiac fibrosis after myocardial infarction impairs cardiac function significantly, pulmonary fibrosis reduces gas exchange efficiency, and liver fibrosis disturbs the natural function of the liver. Its development is associated with the differentiation of fibroblasts into myofibroblasts and increased collagen synthesis. Fibrosis has organ specificity, defined by the heterogeneity of fibroblasts. Although this heterogeneity is established during embryonic development, it has not been defined yet. Fibroblastic differentiation of induced pluripotent stem cells (iPSCs) recapitulates the process by which fibroblasts acquire diversity. Here, we differentiated iPSCs into cardiac, hepatic, and dermal fibroblasts and analyzed their properties using single-cell RNA sequencing. We observed characteristic subpopulations with different ratios in each organ-type fibroblast group, which contained both resting and distinct ACTA2+ myofibroblasts. These findings provide crucial information on the ontogeny-based heterogeneity of fibroblasts, leading to the development of therapeutic strategies to control fibrosis.

Anatomical study of the modified direct posterior gluteal splitting approach for acetabular fractures.

BACKGROUND: In this study, we investigated the area that can be addressed with an approach in which the skin incision is made directly above the dorsal column with Thiel cadaveric specimens. METHODS: Six Thiel cadaveric specimens were prepared. A skin incision was made directly above the dorsal column. The accessible proximal end from the proximal part of the greater sciatic notch to the gluteal ridge and the accessible distal end of the ischium were marked with a flat chisel. A molded 8-hole reconstruction plate was placed from the base of the ischium toward the gluteal ridge and fixed with 3 screws proximally and 2 screws distally. The length of the skin incision and the distance from each reference point on the bone to the reachable markings were assessed after the muscles were removed. RESULTS: Mean skin incision length was 9.3 ± 0.7 (range, 8.0-10.0) cm. In 3 of 6 cases, proximal screws were inserted through different spaces between muscle fibers. In all cases, we were able to reach at least the greater sciatic notch, the gluteal ridge at the level of superior border of the acetabulum, and the base of the ischial tuberosity. In all cases, an 8-hole plate could be placed from the gluteal ridge to the base of the ischium. There were no superior gluteal artery or sciatic nerve injuries in any of the cases. CONCLUSION: We anatomically investigated the area that can be addressed with an approach in which the skin incision was made directly above the dorsal column. In all cases, we were able to access the areas needed to reduce the fracture and place the plates necessary to stabilize the fracture through a 9.3 ± 0.7 cm skin incision. This approach can be a useful minimally invasive posterior approach for acetabular fractures.

The Impact of Potassium Dynamics on Cardiomyocyte Beating in Hemodialysis Treatment.

Background: Observational studies of intermittent hemodialysis therapy have reported that the excess decrease in K+ concentration in plasma (KP) during treatment is associated with the destabilization of cardiac function. Elucidating the mechanism by which the decrease in KP impairs myocardial excitation is indispensable for a deeper understanding of prescription design. Methods: In this study, by using an electrophysiological mathematical model, we investigated the relationship between KP dynamics and cardiomyocyte excitability for the first time. Results: The excess decrease in KP during treatment destabilized cardiomyocyte excitability through the following events: (1) a decrease in KP led to the prolongation of the depolarization phase of ventricular cells due to the reduced potassium efflux rate of the Kr channel, temporarily enhancing contraction force; (2) an excess decrease in KP activated the transport of K+ and Na+ through the funny channel in sinoatrial nodal cells, disrupting automaticity; (3) the excess decrease in KP also resulted in a significant decrease in the resting membrane potential of ventricular cells, causing contractile dysfunction. Avoiding an excess decrease in KP during treatment contributed to the maintenance of cardiomyocyte excitability. Conclusions: The results of these mathematical analyses showed that it is necessary to implement personal prescription or optimal control of K+ concentration in dialysis fluid based on predialysis KP from the perspective of regulatory science in dialysis treatment.

Diafilters for predilution and postdilution on-line hemodiafiltration.

When hemodiafiltration (HDF) is performed, we can choose the point on the blood circuit where the substitution fluid goes in, i.e. before the diafilter (predilution) or after the diafilter (postdilution). We must avoid unexpected loss of albumin; however, we should remove a certain amount of albumin in order to remove much larger solutes than ß2-microglobulin and obtain better clinical outcomes. Previously, ultrafiltration experiments were performed in vitro using aqueous albumin solution and commercial diafilters. The results showed a high sieving coefficient for albumin right after starting the experiment that is identical to several clinical reports. Based on these results, a novel design of the diafilter was attempted for predilution and postdilution HDF. Diafilters for predilution HDF do not require much change in design specifications, employing hollow fibers with a slightly larger diameter in order to reduce high blood pressure caused by the high blood flow rate. Diafilters for postdilution HDF may require much change, including shortening the length in order to reduce the amount of internal filtration. Diafilters specifically designed for predilution or postdilution may be necessary to assure future success of clinical HDF treatment.

A practical design equation for accurate quantification of CRRT filter design factors and convection effects.

Considering the increasing clinical need for continuous renal replacement therapy (CRRT), further improvement in therapeutic efficacy has become an important focus for researchers. Here, we designed nine CRRT filters with various combinations of hollow fiber packing density (PD) and housing shape (effective hollow fiber length (L) and inner housing diameter (D) ratio (L/D ratio)) to evaluate the clearance of middle molecular uremic toxins (MMs) via simulation of an in vitro continuous veno-venous hemodialysis treatment model. We also used Doppler ultrasonography to measure the maximum internal filtration flow rate (QIF-Max) as an aid; this approach facilitated an exploration of the impacts of various design factors on convection effects, while revealing the mechanisms influencing MM removal performance. Furthermore, we constructed a multiple linear regression model of design factors and QIF-Max, then conducted experimental verification. Finally, we proposed an accurate and practical design equation to quantify the design factors influencing CRRT filters and convection effects: QIF-Max=4.749×ND2+2.293×LD-34.775, where N/D2 and L/D affect QIF-Max by 15.0% and 85.0%, respectively. This design equation was able to effectively quantify the convection effects of CRRT filters with different design factors, thereby predicting MM removal performance; this convenient design equation can support the development of CRRT-related products.

Effects of hollow fiber packing density and housing shape on the albumin filtration performance of CRRT filters.

Continuous renal replacement therapy (CRRT) has become the most commonly used acute blood purification therapy for critically ill patients. As a key point of extracorporeal blood circulation, the CRRT filter plays a decisive role in therapeutic efficacy. However, few in vitro studies have been conducted on CRRT filters, particularly concerning the effects of design factors on filter effectiveness and safety profile; no comprehensive evaluation system has been established. Here, we designed nine CRRT filters with various combinations of hollow fiber packing density (PD) and housing shape (effective hollow fiber length (L) and inner housing diameter (D) ratio (L/D ratio)) and introduced a high-frequency sampling pressure monitor to accurately monitor small changes in transmembrane pressure (TMP) and ultrafiltration rate (UFR) over time. We also used concentration polarization mass transfer resistance (Rc), change in sieving coefficient (S) of albumin over time, and amount of albumin removed (Mfld) to investigate the effects of two design factors on albumin filtration performance and analyze the mechanism of protein filtration performance over time, thereby establishing a comprehensive in vitro evaluation system to explore the safety profile of CRRT filters. Our results showed that the nine CRRT filters designed with different combinations of PD (50%, 55%, and 60%) and L/D ratio (2.9, 5.3, and 9.3) were able to maintain stability in terms of hemodynamics and water permeability; the lowest Mfld was PD = 60% and L/D ratio = 9.3, which indicates that design factor optimization can effectively control albumin filtration, thereby improving the safety profile of CRRT filters.