Reduced Mn uptake of pleiotropic ZIP8 SNP is caused by its loss of Mn-responsive accumulation on the cell-surface.

Zrt/Irt-like protein 8 (ZIP8), which is a zinc transporter, plays a pivotal role as a manganese transporter. Recent studies have shown that a ZIP8 SNP (rs13107325 C→T, A391T) is associated with multiple diseases, likely by causing systemic Mn deficiency. However, the underlying molecular mechanisms remain unclear. We attempted to address this issue in cell-based experiments using Madin-Darby canine kidney cells stably expressing ZIP8 WT or the A391T SNP mutant under the control of the Tet-regulatable promoter. We showed that the A391T mutant lost the property of Mn-responsive accumulation on the cell surface, which was observed in WT ZIP8. We also showed that the loss of Mn-responsive accumulation of A391T mutant was associated with its reduced Mn uptake, compared to WT ZIP8, in the Mn uptake assay using the radioisotope 54Mn. Our results potentially explain how the ZIP8 A391T substitution is associated with disease pathogenesis caused by Mn deficiency.

Effects of individual amino acid mutations of zinc transporter ZIP8 on manganese- and cadmium-transporting activity.

Zinc (Zn) transporter ZIP8, encoded by SLC39A8, is a unique transporter that can transport divalent manganese (Mn) and cadmium (Cd) in addition to Zn. Recently, associations between various human diseases and variant forms of ZIP8 have been reported. Four amino acid residues, V33, G38, S335, and I340, of human ZIP8 (hZIP8) are mutated in patients with congenital disorders of glycosylation (CDG), whose blood Mn levels are extremely low. Many genome-wide association studies have reported that the A391T mutation of hZIP8 caused by rs13107325 is associated with a wide range of diseases. However, the roles of individual mutations of hZIP8 on metal-transporting activity remain elusive. We established DT40 cells respectively expressing the four mutant hZIP8s and compared the Mn- and Cd-transporting activity between the mutants and wild-type hZIP8. Among the four mutations observed in the ZIP8-mutated CDG patients, the S335T and I340 N mutations in the predicted transmembrane domain 5 (TMD5) completely abolished Mn- and Cd-transporting activity, while V33 M or G35R mutations at the N-terminus did not. We also examined the A391T mutation, which slightly reduced metal transporting activity. Finally, we examined the effects of artificial mutations in the metal-binding motif EEXXH in the TMD5. Replacing EEXXH with HEXXH, which exists in most ZIP transporters, abolished the Mn- and Cd-transporting activity of hZIP8, indicating that glutamic acid in this motif plays a critical role in the unique affinity of ZIP8 for Mn and Cd. Thus, the utilization of DT40 cells enabled us to clarify the different functions of each residue of hZIP8 on metal transport.

Manganese transport in mammals by zinc transporter family proteins, ZNT and ZIP.

Manganese (Mn) is an essential trace element required for various biological processes. However, excess Mn causes serious side effects in humans, including parkinsonism. Thus, elucidation of Mn homeostasis at the systemic, cellular, and molecular levels is important. Many metal transporters and channels can be involved in the transport and homeostasis of Mn, and an increasing body of evidence shows that several zinc (Zn) transporters belonging to the ZIP and ZNT families, specifically, ZNT10, ZIP8, and ZIP14, play pivotal roles in Mn metabolism. Mutations in the genes encoding these transporter proteins are associated with congenital disorders related to dysregulated Mn homeostasis in humans. Moreover, single nucleotide polymorphisms of ZIP8 are associated with multiple clinical phenotypes. In this review, we discuss the recent literature on the structural and biochemical features of ZNT10, ZIP8, and ZIP14, including transport mechanisms, regulation of expression, and pathophysiological functions. Because a disturbance in Mn homeostasis is closely associated with a variety of phenotypes and risk of human diseases, these transporters constitute a significant target for drug development. An understanding of the roles of these key transporters in Mn metabolism should provide new insights into pharmacological applications of their inhibitors and enhancers in human diseases.

New Insights into the Roles of ZIP8, a Cadmium and Manganese Transporter, and Its Relation to Human Diseases.

ZIP8, a Zrt-/Irt-related protein encoded by Slc39A8, was originally discovered as a zinc transporter, but since then its roles as a transporter for cadmium (Cd) and manganese (Mn) have also been well characterized. ZIP8 is highly expressed in the S3 segment of the proximal tubules of the mouse kidney and may play a significant role in reabsorption of both toxic Cd and essential Mn from the lumen to the epithelial cells of the proximal tubule. In recent years, associations between various human diseases and genetic variations of ZIP8 have been reported. Missense mutations in the human SLC39A8 gene are associated with serious disorders of Mn metabolism, showing symptoms similar to congenital glycosylation deficiency. Enhanced excretion of Mn via bile or urine might be the cause of extremely low blood Mn levels in ZIP8-mutated patients, leading to the defects in Mn-dependent glycosylation. Several genome-wide association studies have demonstrated the associations of multiple diseases and ZIP8 SNPs constituting missense mutations. These findings suggest that ZIP8 plays more important roles than previously expected as a modulator of Mn homeostasis in the body. Elucidation of biochemical mechanisms regarding the metal-transporting ability of ZIP8 and its alteration by mutation is required for better understanding of the role of ZIP8 in human diseases.

Protective effect of cadmium-induced autophagy in rat renal mesangial cells.

Cadmium damages renal cells, and in particular may cause mesangial cell death by necrosis or apoptosis, depending on exposure conditions in cultured cells. However, there is an uncertainty as to whether Cd2+-induced autophagy can protect mesangial cells against these other mechanisms of cell death. We have used autophagy-incompetent mouse embryonic fibroblast (MEF) cells lacking the Atg16 gene, as well as cultured rat mesangial cells (RMC) in which Atg16 has been silenced, to examine this issue. Measuring the processing of LC3-I to LC3-II and expression of sequestosome-1 (p62), we define conditions under which RMC can be induced to undergo autophagy in response to 0-20 µM CdCl2. Similarly, Cd2+ can initiate autophagy in MEF cells. However, when autophagy is compromised, either by gene knockout in MEF cells or by RNA silencing in RMC, cell viability is decreased, and concomitantly a Cd2+ dose-dependent increase in pro-caspase-3 cleavage indicates the initiation of apoptotic cell death. In contrast to some previous reports, Cd2+-induced autophagy is not correlated with increased levels of cellular reactive oxygen species but, among a panel of kinases investigated, is suppressed by inhibition of the Jun kinase. We conclude that concentrations of Cd2+ that initiate autophagy may afford renal mesangial cells some degree of protection against other modes (apoptosis, necrosis) of cell death.

An anatomic study on the attachment of the joint capsule to the tibia in the lateral side of the knee.

PURPOSE: The purpose of the current study was to examine the width, area, and histological characteristics of the capsular attachment to the tibia in the lateral side of the knee. METHODS: A total of 27 knees were used in this study. The joint capsule of the knee was peeled away from the tibia and the width of the capsular attachment to the tibia was measured by two independent observers using a caliper. Interclass correlation coefficients for each value were calculated to evaluate the validity of the measurement. The capsular attachment to the tibia of the seven knees was histologically analyzed using Masson's trichrome staining. RESULTS: At the posterior border of Gerdy's tubercle, the capsular attachment was wide; the average width was 8.6 mm (SD 3.0). Toward the posterolateral aspect of the knee, the capsular attachment gradually tapered. Finally, the capsular attachment was linear at the apex of the head of the fibula. Histological analysis at the posterior border of Gerdy's tubercle revealed developed uncalcified fibrocartilage on the capsular attachment. In contrast, at the apex of the head of the fibula, the joint capsule was adhered to the capsule of the proximal tibiofibular joint. Fibrous connective tissue was directly attached to the calcified fibrocartilage. CONCLUSIONS: The attachment width of the knee joint capsule at the lateral side varied according to location. We consider that this finding on the capsular attachment will facilitate an understanding of the pathology or mechanism of diseases on the lateral side of the knee joint.

Direct Comparison of Manganese Detoxification/Efflux Proteins and Molecular Characterization of ZnT10 Protein as a Manganese Transporter.

Manganese homeostasis involves coordinated regulation of specific proteins involved in manganese influx and efflux. However, the proteins that are involved in detoxification/efflux have not been completely resolved nor has the basis by which they select their metal substrate. Here, we compared six proteins, which were reported to be involved in manganese detoxification/efflux, by evaluating their ability to reduce manganese toxicity in chicken DT40 cells, finding that human ZnT10 (hZnT10) was the most significant contributor. A domain swapping and substitution analysis between hZnT10 and the zinc-specific transporter hZnT1 showed that residue Asn(43), which corresponds to the His residue constituting the potential intramembranous zinc coordination site in other ZnT transporters, is necessary to impart hZnT10's unique manganese mobilization activity; residues Cys(52) and Leu(242) in transmembrane domains II and V play a subtler role in controlling the metal specificity of hZnT10. Interestingly, the His → Asn reversion mutant in hZnT1 conferred manganese transport activity and loss of zinc transport activity. These results provide important information about manganese detoxification/efflux mechanisms in vertebrate cells as well as the molecular characterization of hZnT10 as a manganese transporter.

Zinc transporter SLC39A10/ZIP10 controls humoral immunity by modulating B-cell receptor signal strength.

The humoral immune response, also called the antibody-mediated immune response, is one of the main adaptive immune systems. The essential micronutrient zinc (Zn) is known to modulate adaptive immune responses, and dysregulated Zn homeostasis leads to immunodeficiency. However, the molecular mechanisms underlying this Zn-mediated modulation are largely unknown. Here, we show that the Zn transporter SLC39A10/ZIP10 plays an important role in B-cell antigen receptor (BCR) signal transduction. Zip10-deficiency in mature B cells attenuated both T-cell-dependent and -independent immune responses in vivo. The Zip10-deficient mature B cells proliferated poorly in response to BCR cross-linking, as a result of dysregulated BCR signaling. The perturbed signaling was found to be triggered by a reduction in CD45R phosphatase activity and consequent hyperactivation of LYN, an essential protein kinase in BCR signaling. Our data suggest that ZIP10 functions as a positive regulator of CD45R to modulate the BCR signal strength, thereby setting a threshold for BCR signaling in humoral immune responses.

Attachment area of fibres from the horns of lateral meniscus: anatomic study with special reference to the positional relationship of anterior cruciate ligament.

PURPOSE: Although studies support the clinical importance of the fibres from the horns of lateral meniscus (LM), few studies have investigated the detailed anatomy. This anatomic study was conducted to analyse the structural details of LM with special reference to (1) the attachment area of the fibres from the anterior and posterior horns and (2) the positional relationship between these fibres and the anterior cruciate ligament (ACL). METHODS: A total of 24 cadaveric knees were used in the macroscopic investigation, and six knees were used in the histological investigation. Micro-computed tomography analysis was also performed to assess the anatomy of the posteriormost fibre from the posterior horn of LM. RESULTS: Based on the macroscopic investigations, the outer fibres from the anterior horn of LM extended to ACL and seemed to intermingle with ACL fibres. However, the histological investigations showed a distinct border between the fibres and ACL. The inner fibres from the anterior horn of LM attached to the lateral intercondylar tubercle serving as a lateral margin of ACL attachment. Fibres from the posterior horn of LM were separated into anterolateral and posteromedial crura which attached to the posterior aspect of the lateral and medial intercondylar tubercles, respectively. These two crura formed the posterior margin of the ACL attachment, except for the central part of ACL. CONCLUSION: The outer fibres from the anterior horn of LM adjoined ACL. The inner fibres from the anterior horn of LM and two crura from the posterior horn of LM formed the border of the attachment area of ACL. The distinctive fibre anatomy from LM could provide a surgical landmark during arthroscopic surgery.

Joint capsule attachment to the coronoid process of the ulna: an anatomic study with implications regarding the type 1 fractures of the coronoid process of the O'Driscoll classification.

BACKGROUND: The attachment of the anterior joint capsule on the ulnar coronoid process is not yet completely understood. The purpose of this study was to clarify the anatomic relationship between the anterior capsule of the elbow joint and the tip of the coronoid process. METHODS: Seventeen embalmed elbows were used for this anatomic study. The anterior capsule of the elbow joint was reflected, and the attachment of the capsule on the coronoid process was exposed. The attachment of the joint capsule on the coronoid process was macroscopically and histologically observed, its relationship to the coronoid tip was assessed, and the length of the attachment of the joint capsule was measured. RESULTS: The length of the capsule attachment at the radial side of the coronoid (11.9 mm) was greater than that at the ulnar side (6.1 mm). The bone thickness on the coronoid tip from the proximal edge of the joint capsule attachment was 1.9 mm; together, the cartilage and bone thickness was 4.7 mm. At the radial side of the coronoid, the thickness of the joint capsule at the proximal aspect of the attachment of 2 samples was 0.6 mm and 0.3 mm, and that at the tip of the coronoid was 2.6 mm and 1.7 mm, respectively. CONCLUSIONS: The anterior capsule of the elbow joint had a substantial attachment on the radial side of the coronoid process. The subtype 2 tip fractures of the O'Driscoll classification included the joint capsule attachment, joint cartilage, and subchondral bone.

An anatomic study of the attachments on the condylar process of the mandible: muscle bundles from the temporalis.

PURPOSE: The aim of this study was to evaluate anatomically the relationship between bone and muscles by detailed observation of the bone shape and the structure of muscles to facilitate an understanding of the function of the muscles involved in jaw movement. METHODS: 36 specimens of 24 Japanese cadavers were examined. The insertion areas were marked using a radiopaque marker and examined by micro-computed tomography. For morphological observation, we used 101 condylar processes. In addition, we made histological sections in some specimens to observe the detailed attachments of the muscle. RESULTS: Based on the micro-CT images and dissection findings, the lateral pterygoid muscle was found to be most frequently inserted into the anterior impression and attached to the medial impression of the process. According to the histological observations, the lateral pterygoid muscle mainly inserted to the condylar process. The micro-CT images indicated that the obvious bony ridge was lateral to the pterygoid fovea on the condylar process in all specimens. The midmedial muscle bundle of the temporalis was attached to the ridge. Based on the morphological observations, the ridge was situated on the lateral area of the condylar process. CONCLUSIONS: Since dysfunction of the temporomandibular joint is likely closely related to both the lateral pterygoid muscle and also the temporalis, further studies are necessary to evaluate the function of these muscles and consider jaw movement.

The anatomic relationship between the morphology of the greater tubercle of the humerus and the insertion of the infraspinatus tendon.

BACKGROUND: The objective of this study was to evaluate the topographic relationship between the morphology of the greater tubercle and the insertion of the tendon of the infraspinatus. MATERIALS AND METHODS: First, we defined an impression of the greater tubercle, which has not been recognized in classic textbooks, as the "lateral impression" and then measured the dimensions of the "lateral impression" of the greater tubercle in 71 samples of dry bone of humeri. Next, we examined 16 cadaveric humeri with rotator cuff tendons by micro-computed tomography to analyze the positional relationship between the lateral impression and the infraspinatus tendon. RESULTS: In all samples of dry bones, the lateral impression could be identified as a triangle shape. The lateral impression was composed of the border with the highest impression (mean, 6.3 mm), the border with the middle impression (mean, 5.0 mm), and the border with the lateral wall of the greater tubercle (mean, 8.5 mm). In all samples of humeri with rotator cuffs, we could confirm the lateral impression, and the border between the highest impression and the lateral impression corresponded to the anterior border of the insertion of the infraspinatus tendon. CONCLUSION: We propose a new anatomic concept of the lateral impression that could enable the precise diagnosis of and facilitate repair techniques for infraspinatus tear, according to specific anatomic characteristics, by applying 3-dimensional computed tomography assessment preoperatively.

Anatomic and histologic analysis of the mid-substance and fan-like extension fibres of the anterior cruciate ligament during knee motion, with special reference to the femoral attachment.

PURPOSE: The purpose of this study is to evaluate the morphology of the mid-substance and fan-like extension fibres of ACL during knee motion with reference to the femoral attachment. METHODS: This study used six fresh-frozen cadaveric knees and 22 embalmed cadaveric knees to macroscopically evaluate morphological changes in the ACL attachment during knee motion. Three embalmed specimens fixed at knee extension and another three specimens fixed at 120° flexion were used for histologic observations. RESULTS: The fan-like extension fibres were adhered to the bone surface and the fibre location and orientation in relation to the femoral surface did not change, regardless of the knee flexion angle, while the orientation of the mid-substance fibres in relation to the femur did change during knee motion. During knee flexion, a fold in the ACL femoral attachment was observed at the border between the mid-substance and the fan-like extension fibres. The attachment of the mid-substance fibres was significantly smaller than the attachment of the fan-like extension fibres. CONCLUSION: The present study clarified anatomic and histologic character of the mid-substance fibres and fan-like extension fibres, and provided critical information for future clinical and biomechanical studies concerning both two different fibres. Specifically for ACL reconstruction, it is difficult to reconstruct the natural fan-like extension fibres by creating a tunnel at the femoral and tibial ends of each fibre bundle, although the mid-substance fibres can be reconstructed by such procedures.

Joint capsule attachment to the extensor carpi radialis brevis origin: an anatomical study with possible implications regarding the etiology of lateral epicondylitis.

PURPOSE: To identify the unique anatomical characteristic of the extensor carpi radialis brevis (ECRB) origin and points of differentiation from other extensors and to clarify the specific relationship of the ECRB to the underlying structures. METHODS: We studied the origin of each extensor macroscopically for its muscular and tendinous parts; to identify the relationship between the ECRB origin and the deeper structures, we also examined the attachment of the joint capsule under the ECRB origin. RESULTS: The ECRB simply originated as a tendon without any muscle, whereas other extensors originated as a mixture of tendon and muscle. At the anterior part of the ECRB origin, the thin attachment of the joint capsule (average width, 3.3 mm) lay deep to the ECRB and was distinct. However, at the posterodistal portion, the joint capsule, annular ligament, and supinator were intermingled and originated as a single wide sheet from the humerus (average width, 10.7 mm). CONCLUSIONS: The anterior part of the ECRB origin was delicate, because the ECRB origin was purely tendinous, and the attachment of the articular capsule was thin compared with that of the posterodistal attachment. This thin attachment could be an initial factor leading to the development of lateral epicondylitis. CLINICAL RELEVANCE: The results of the current study may enhance magnetic resonance imaging understanding and may help clarify the etiology of the lateral epicondylitis.

Ferroptosis is involved in cisplatin sensitivity of the S3 segment of immortalized proximal tubule cells.

Cisplatin (CDDP) is administered as an anticancer drug across a broad spectrum of cancer treatments, but it causes severe renal damage. Several studies have attempted to elucidate the cause of CDDP-induced renal injury, but the detailed mechanism remains unclear. We previously found that S3 cells are more sensitive to CDDP than S1 and S2 cells by using immortalized cells derived from S1, S2, and S3 segments of proximal tubules. In this study, we investigated the potential contribution of reactive oxygen species (ROS) to the sensitivity of S3 cells to CDDP. The results showed that S3 cells have high sensitivity to CDDP, paraquat (PQ) and three ROS substances. To examine the mechanisms underlying the sensitivity to ROS in S3 cells, we compared the cellular responses of CDDP- and PQ-exposed S3 cells. The results indicated that the levels of intracellular ROS and lipid peroxides were increased in S3 cells after CDDP and PQ exposure. The intracellular levels of antioxidant proteins such as thioredoxin, thioredoxin reductase 1 and glutathione peroxidase 4 were also increased by exposure to PQ, but these proteins were decreased by CDDP exposure in S3 cells. Furthermore, the levels of intracellular free Fe2+ were increased by CDDP exposure only in S3 cells but not S1 or S2 cells, and cytotoxicity by exposure to CDDP in S3 cells was suppressed by ferroptosis inhibitors. These results suggested that the induction of ferroptosis due to the ROS production through attenuation of the antioxidant system and elevated free Fe2+ is partly responsible for the sensitivity of S3 cells to CDDP.

CHAC1 exacerbates arsenite cytotoxicity by lowering intracellular glutathione levels.

We here examined whether CHAC1 is implicated in arsenite (As(III))-induced cytotoxicity in HaCaT cells. We found that HaCaT cells in which the intracellular GSH levels were elevated by transfection with CHAC1 siRNA showed decreased sensitivity to As(III) compared to the control cells. Treatment with BSO (an inhibitor of GSH biosynthesis) abolished the decrease in sensitivity to As(III), suggesting that an increase in intracellular GSH levels was involved in the decrease in sensitivity to As(III) due to the decrease in the levels of CHAC1 expression. When we examined the expression of CHAC1 after exposure of HaCaT cells to As(III), the levels of CHAC1 were increased. Since CHAC1 is a proapoptotic factor, we examined appearance of apoptotic cells and cleavage of caspase-3 after exposure to As(III) to determine whether As(III)-induced CHAC1 up-regulation was involved in apoptosis induction. The results showed that induction of apoptosis by As(III) exposure was not detected in CHAC1 siRNA-transfected cells. Together, our findings indicate that CHAC1 is involved in the sensitivity of HaCaT cells to As(III) by regulating the intracellular GSH levels, and in particular, CHAC1 is involved in As(III)-induced apoptosis.

Anatomical study of the bone morphology of the anterior talofibular ligament attachment.

Anterior talofibular ligament (ATFL) injuries are the most common cause of ankle sprains. To ensure anatomically accurate surgery and ultrasound imaging of the ATFL, anatomical knowledge of the bony landmarks around the ATFL attachment to the distal fibula is required. The purpose of the present study was to anatomically investigate the ATFL attachment to the fibula with respect to bone morphology and attachment structures. First, we analyzed 36 feet using microcomputed tomography. After excluding 9 feet for deformities, the remaining 27 feet were used for chemically debrided bone analysis and macroscopic and histological observations. Ten feet of living specimens were observed using ultrasonography. We found that a bony ridge was present at the boundary between the attachments of the ATFL and calcaneofibular ligament (CFL) to the fibula. These two attachments could be distinguished based on a difference in fiber orientation. Histologically, the ATFL was attached to the anterodistal part of the fibula via fibrocartilage anterior to the bony ridge indicating the border with the CFL attachment. Using ultrasonography in living specimens, the bony ridge and hyperechoic fibrillar pattern of the ATFL could be visualized. We established that the bony ridge corresponded to the posterior margin of the ATFL attachment itself. The ridge was obvious, and the superior fibers of the ATFL have directly attached anteriorly to it. This bony ridge could become a valuable and easy-to-use landmark for ultrasound imaging of the ATFL attachment if combined with the identification of the fibrillar pattern of the ATFL.

Effects of methylation of arginine residue 83 on the enzymatic activity of human arsenic (+3 oxidation state) methyltransferase.

Arsenic (+3 oxidation state) methyltransferase is an enzyme responsible for arsenic methylation, and it requires S-adenosyl-methionine (SAM) as a coenzyme. We here generated two mutants to clarify the role of the highly conserved 83rd arginine residue (Arg83) in Motif I, the SAM-binding domain, of human AS3MT. When the AS3MT activity was compared between the mutants and the wild type (WT) recombinant protein, little activity was detected in the glycine mutant (Arg83Gly) or lysine mutant (Arg83Lys). When we examined the ability of transfected HEK293 cells exposed to arsenite to methylate arsenic, the methylation ability was significantly reduced in Arg83Gly compared to the WT, but was not significantly different between Arg83Lys and WT. Western blot analysis of the recombinant WT and Arg83Gly with an antibody that recognizes methylated Arg showed that an Arg residue in the WT was mono- and di-methylated, but not in Arg83Gly. Furthermore, a peptide containing dimethylated Arg83 was detected by MALDI-TOF/MS of the WT digested with chymotrypsin. These results indicate that AS3MT maintains its enzymatic activity through the methyl modification of Arg83.

Lateral Ulnar Collateral Ligament of the Elbow Joint: Reconsideration of Anatomy in Terms of Connection with Surrounding Fibrous Structures.

BACKGROUND: To improve the clinical results of lateral ulnar collateral ligament (LUCL) reconstruction of the elbow joint, better understanding of the anatomy of the aponeuroses and joint capsule could be relevant. This study considers the previously described anatomy of the LUCL in relation to the related aponeuroses and joint capsule rather than as a discrete ligament. We hypothesized that the deep aponeuroses of the superficial extensor muscles and supinator form a relevant portion of the joint capsule previously defined as the LUCL. METHODS: Twenty-four elbows (12 right) from 21 embalmed cadavers (age at the time of death, 54 to 99 years) were included in the study. Twenty elbows were studied macroscopically and 4, histologically. The joint capsule was detached from the bones, and local thickness was quantitatively analyzed using micro-computed tomography (micro-CT). RESULTS: The supinator aponeurosis and joint capsule intermingled to form a thick membrane (mean and standard deviation, 4.8 ± 1.2 mm), which we termed "the capsulo-aponeurotic membrane." It was thicker than the anterior (1.3 ± 0.4 mm) and posterior (2.5 ± 0.9 mm) parts of the capsule of the humeroradial joint (p < 0.001). The capsulo-aponeurotic membrane had a wide attachment on the distal part of the extensor digitorum communis and extensor digiti minimi (EDC/EDM) origin of the humerus, the lateral part of the coronoid process, and the posterior part of the radial notch of the ulna. The humeral attachment had a fibrocartilaginous structure. The deep aponeuroses of the EDC and extensor carpi ulnaris (ECU) were connected to the capsulo-aponeurotic membrane. CONCLUSIONS: The capsulo-aponeurotic membrane was composed of the supinator aponeurosis and joint capsule and was attached to the lateral epicondyle of the humerus, radial side of the coronoid process, and posterior part of the radial notch on the ulna. The entire structure appeared identical to the commonly defined lateral collateral ligament. The most posterior part was connected to the EDC and ECU aponeuroses, which is commonly labeled the LUCL but does not exist as a discrete ligament. CLINICAL RELEVANCE: Consideration of the accurate anatomy of the extensive attachment of the capsulo-aponeurotic membrane could provide useful clues for improvement in techniques of LUCL reconstruction and lateral epicondylitis pathology.

Spatial localization of cadmium and metallothionein in the kidneys of mice at the early phase of cadmium accumulation.

Chronic exposure to cadmium (Cd) leads to an accumulation of Cd in the kidneys. Metallothionein (MT) is a low-molecular-weight protein having a high affinity for Cd. Cd bound to MT in serum is filtered through the glomeruli of kidney nephrons and reabsorbed by endocytosis into the proximal tubules from the luminal side. Accumulation of Cd in renal cells induces MT synthesis, leading to long-term deposition of Cd and the suppression of Cd toxicity. Recently, many studies have investigated the tissue distribution of metals using laser ablation ICP-MS (LA-ICP-MS). However, little information has been available regarding renal Cd distribution. Hence, we dually investigated the renal distribution of Cd by LA-ICP-MS and that of MT by immunohistochemistry to clarify the dose- and time-dependent changes in the distributions of Cd and MT in mice exposed to Cd from drinking water for 1, 2, and 4 months. Both Cd and MT exhibited characteristic heterogeneous distribution patterns in the renal cortex. The accumulation of Cd and MT near the surface of the cortex suggests a preferential accumulation of Cd in the surface nephrons. MT distribution was more pronounced in the proximal tubules than in the distal tubules, and there were clear differences in MT immunostaining even among the proximal tubules. The heterogeneous localization of MT may reflect the nephron-specific accumulation of Cd. Combining elemental imaging of Cd with immunostaining of MT proved a successful strategy to reveal the characteristic renal Cd distribution, especially in the early stages of Cd accumulation.