Short-term results and surgical technique of arthroscopic centralization as an augmentation for medial meniscus extrusion caused by medial meniscus posterior root tear.

INTRODUCTION: The purpose of this study was to perform medial meniscus (MM) centralization for medial meniscus extrusion (MME) associated with medial meniscus posterior root tear (MMPRT) and to examine the short-term results. The hypothesis is that arthroscopic centralization as an augmentation of MMPRT repair improves clinical outcomes and the extrusion distance of MM in short-term results. MATERIALS AND METHODS: Twenty-six patients (mean age 62.1 ± 6.0 years) who underwent arthroscopic centralization as an augmentation of MMPRT repair were included. Clinical evaluation was performed before and 2 years after surgery using Lysholm score and knee injury and osteoarthritis outcome score (KOOS). Image evaluation used MRI and plain X-ray images. The extrusion distance and MME ratio were compared on MRI images before and 2 years after surgery. The degree of osteoarthritis (OA) was evaluated using Kellgren-Lawrence classification. The degree of OA and hip-knee-ankle (HKA) angle were compared by plane X-ray images before and 2 years after surgery. RESULTS: In clinical results, both Lysholm score and KOOS improved significantly after surgery. In image evaluation, the extrusion distance decreased significantly from 4.8 ± 0.7 mm before surgery to 2.7 ± 0.3 mm 2 years after surgery (p < 0.05). The MME ratio was significantly improved from 40.2 ± 7.0% before surgery to 22.6 ± 3.6% after surgery (p < 0.05). There was no significant difference in HKA angle at 2 years after surgery (p = 0.13). CONCLUSIONS: The arthroscopic centralization for medial meniscal extrusion associated with MMPRT significantly improved clinical outcomes and the extrusion distance of MM. It is also one of the surgical techniques that can suppress medial meniscus extrusion. LEVEL OF EVIDENCE: IV, therapeutic case series.

Arthroscopic ankle lateral ligament repair alone versus arthroscopic ankle lateral ligament repair with reinforcement by inferior extensor retinaculum.

PURPOSE: This study aimed to compare the clinical outcomes and postoperative activities of arthroscopic ankle lateral ligament (ALL) repair alone with arthroscopic ALL repair and reinforcement by the inferior extensor retinaculum (IER) for chronic ankle instability (CAI). MATERIALS AND METHODS: All patients who underwent arthroscopic repair for CAI between 2017 and 2019 were evaluated. The Japanese Society for Surgery of the Foot (JSSF) scale and self-administered foot evaluation questionnaire (SAFE-Q), and duration between the surgery and walking without any support, jogging, and complete return to sports were evaluated and compared. The exclusion criteria were (1) follow-up period of < 1 year after surgery, (2) the presence of associated ankle lesions requiring treatment during the same operative procedure, including patients with subfibular ossicle bigger than 5 mm on radiographs, chondral or osteochondral defect, bony impingement, deltoid ligament tear, fibular tendon pathology, or posterior ankle impingement, and (3) patients who underwent revision surgery. RESULTS: We identified 126 patients who underwent surgery for CAI and subsequently excluded 36 patients on account of a short follow-up period (< 1 year), additional surgery, and previous surgery. The remaining 90 eligible patients included arthroscopic ALL repair alone (group A, n = 44) and arthroscopic ALL repair with reinforcement by the inferior extensor retinaculum (group G, n = 46) groups. There was no significant difference in the postoperative activities nor in the preoperative or postoperative JSSF scale and SAFE-Q between the two groups. However, significant differences were seen in the mean surgical time (15.5 ± 8.1 vs 20.1 ± 7.6, P = 0.013). CONCLUSION: This study showed no difference in clinical outcomes between the two groups. However, arthroscopic ALL repair with reinforcement by IER resulted in a longer surgical time than arthroscopic ALL repair alone. LEVEL OF EVIDENCE: Retrospective comparative study, level III.

Subunit structure and inhibition specificity of alpha-type phospholipase A2 inhibitor from Protobothrops flavoviridis.

The alpha-type phospholipase A2 inhibitor (PLIalpha) in the plasma of the Habu snake, Protobothrop flavoviridis, was shown to be a trimer of two homologous subunits, PLIalpha-A and PLIalpha-B, each of which contains one C-type lectin-like domain (CTLD). Since one molecule of trimeric PLIalpha binds stoichiometrically to one molecule of P. flavoviridis acidic phospholipase A2 (PLA2), the trimeric structure is critical for its inhibitory activity. Hydrophobic chromatography separated the purified P. flavoviridis PLIalpha into four different trimeric subspecies, A3-PLIalpha, A2B-PLIalpha, AB2-PLIalpha, and B3-PLIalpha, with different combinations of the two subunits. The trimeric PLIalpha could be reconstituted from the purified subunits, and the four different trimeric subspecies were formed through random association of the two subunits. The inhibitory activity of the PLIalpha-A homotrimer (A3-PLIalpha) was more specific than that of the PLIalpha-B homotrimer (B3-PLIalpha). This difference in inhibitory properties between the two homotrimers was probably caused by the amino acid differences at residues 10-37.

Structural analysis of the interaction between Shiga toxin B subunits and linear polymers bearing clustered globotriose residues.

We previously developed linear polymers bearing clustered trisaccharides of globotriaosylceramide (Gb3) as orally applicable Shiga toxin (Stx) neutralizers. Here, using a Gb3 polymer with a short spacer tethering the trisaccharide to the core, we found that shortening the spacer length markedly reduced the binding affinity for Stx2 but not Stx1. Moreover, mutational analysis revealed that the essential binding sites of the terminal trisaccharides were completely different between Stx1 and Stx2. These results provide the molecular basis for the interaction between Stx B subunits and Gb3 polymers.

Molecular cloning of the major lethal toxins from two kraits (Bungarus flaviceps and Bungarus candidus).

The major lethal toxins present in the venoms of the red-headed krait, Bungarus flaviceps, and the Malayan krait, Bungarus candidus, have both been purified. Each consists of two polypeptide chains, A and B, joined by a disulfide bond. In the present study, primary structures of these toxins were determined by Edman degradation and by nucleotide sequencing of the cDNA clones. Amino acid sequencing of the N-terminus and enzymatically digested peptides revealed that the A and B chains were highly homologous to those of beta-bungarotoxins (beta-Bgts) from Bungarus multicinctus, respectively. We isolated cDNA clones encoding the A and B chains from both B. flaviceps and B. candidus venom gland cDNA libraries using probes designed based on the cDNA sequence of beta-Bgt from B. multicinctus. Two isoforms of the A chain and one isoform of the B chain were obtained from B. flaviceps, and one isoform of the A chain and two isoforms of the B chain were obtained from B. candidus. Both of the two A chains from B. flaviceps are made up of 119 amino acids and comprise 15 cysteine residues, while the A chains of beta-Bgt from other Bungarus species including B. candidus comprise 13 cysteine residues. The B chains from both species are composed of 59 amino acid residues and comprise seven cysteines. In conclusion, the lethal toxin from B. flaviceps is considered to be a novel isoform of beta-Bgt, which has a different pattern of cysteine residues from known beta-Bgts.

Complete amino acid sequence and phylogenetic analysis of a long-chain neurotoxin from the venom of the African banded water cobra, Boulengerina annulata.

The sequence of a long-chain neurotoxin (71 amino acid residues, 10 half-cystines) from the venom of the African banded water cobra (Boulengerina annulata) was determined by Edman degradation. It exhibits high sequence similarity with long-chain neurotoxins from the venoms of four species of African cobras (genus Naja), which are collectively more similar to the Boulengerina toxin than to those of Asian Naja species. These results are discussed in the light of phylogenetic hypotheses on the relationships of African cobras.

Oral therapeutic agents with highly clustered globotriose for treatment of Shiga toxigenic Escherichia coli infections.

Shiga toxin (Stx) is a major virulence factor in infection with Stx-producing Escherichia coli (STEC). We developed a series of linear polymers of acrylamide, each with a different density of trisaccharide of globotriaosylceramide (Gb3), which is a receptor for Stx, and identified Gb3 polymers with highly clustered trisaccharides as Stx adsorbents functioning in the gut. The Gb3 polymers specifically bound to both Stx1 and Stx2 with high affinity and markedly inhibited the cytotoxic activities of these toxins. Oral administration of the Gb3 polymers protected mice after administration of a fatal dose of E. coli O157:H7, even when the polymers were administered after the infection had been established. In these mice, the serum level of Stx was markedly reduced and fatal brain damage was substantially suppressed, which suggests that the Gb3 polymers entrap Stx in the gut and prevent its entrance into the circulation. These results indicate that the Gb3 polymers can be used as oral therapeutic agents that function in the gut against STEC infections.

Identification of alpha-bungarotoxin (A31) as the major postsynaptic neurotoxin, and complete nucleotide identity of a genomic DNA of Bungarus candidus from Java with exons of the Bungarus multicinctus alpha-bungarotoxin (A31) gene.

The Malayan krait (Bungarus candidus) is one of the most medically significant snake species in Southeast Asia. No specific antivenom exists to treat envenoming by this species. Death within 30 min after its bite has been reported from Java, suggesting the presence of highly lethal postsynaptic neurotoxins in the venom of these snakes. We purified and identified the major postsynaptic toxin in the venom of B. candidus from Java. The toxin was indistinguishable from alpha-bungarotoxin (A31), a toxin originally isolated from Bungarus multicinctus, in its mass (7983.75 Da), LD50 (0.23 microg/g in mice i.p.), affinity to nicotinic acetylcholine receptors, and by its 40 N-terminal amino acid residues as determined by Edman degradation. Identity with alpha-bungarotoxin was confirmed by cloning and sequencing a genomic DNA from B. candidus which encodes the 74 amino acid sequence of alpha-bungarotoxin (A31) and part of its signal peptide, revealing complete identity to the alpha-bungarotoxin (A31) gene in exon and 98.9% identity in intron sequences. The entire mitochondrial cytochrome b gene of the krait species B. candidus from Java and B. multicinctus from Taiwan was sequenced for comparison, suggesting that these snakes are phylogenetically closely related. alpha-Bungarotoxin appears to be widely present and conserved in Southeast and East Asian black-and-white kraits across populations and taxa.

Isolation, toxicity and amino terminal sequences of three major neurotoxins in the venom of Malayan krait (Bungarus candidus) from Thailand.

We isolated the most lethal toxins in the venom of the Malayan krait (Bungarus candidus), one of the medically most important snake species in southeast Asia. Three beta-BTx like basic neurotoxins, T1-1, T1-2, and T2, with PLA2 activity were isolated from pooled venom of eight B. candidus from southern Thailand by cation-exchange chromatography, followed by adsorption chromatography on hydroxylapatite and RP-HPLC, with 14-, 16-, and 4-fold increases in toxicity compared to crude venom. The LDs50 determined in mice weighing 18-20 g were 0.26, 0.22, and 0.84 micro g per mouse with i.v. injection. T1-1 and T1-2 possessed comparable lethal toxicities to those of beta1-BTx, the most toxic neurotoxin in B. multicinctus venom, and the major neurotoxin in B. flaviceps venom. The apparent molecular weights of the native toxins were approximately 25-25.5 kDa. They consist of two polypeptide chains with apparent molecular weights of 15.5-16.5 and 8-8.5 kDa, respectively. The amino terminal sequences of the two chains of each of the toxins determined by Edman degradation exhibited considerable similarity with those of the A-chains and B-chains of beta-BTxs in the venom of Bungarus multicinctus.