Recent Progress Toward Hydrogen Medicine: Potential of Molecular Hydrogen for Preventive and Therapeutic Applications

Persistent oxidative stress is one of the major causes of most lifestyle-related diseases, cancer and the aging process. Acute oxidative stress directly causes serious damage to tissues. Despite the clinical importance of oxidative damage, antioxidants have been of limited therapeutic success. We have proposed that molecular hydrogen (H2) has potential as a “novel” antioxidant in preventive and therapeutic applications [Ohsawa et al., Nat Med. 2007: 13; 688-94]. H2 has a number of advantages as a potential antioxidant: H2 rapidly diffuses into tissues and cells, and it is mild enough neither to disturb metabolic redox reactions nor to affect reactive oxygen species (ROS) that function in cell signaling, thereby, there should be little adverse effects of consuming H2. There are several methods to ingest or consume H2, including inhaling hydrogen gas, drinking H2-dissolved water (hydrogen water), taking a hydrogen bath, injecting H2-dissolved saline (hydrogen saline), dropping hydrogen saline onto the eye, and increasing the production of intestinal H2 by bacteria. Since the publication of the first H2 paper in Nature Medicine in 2007, the biological effects of H2 have been confirmed by the publication of more than 38 diseases, physiological states and clinical tests in leading biological/medical journals, and several groups have started clinical examinations. Moreover, H2 shows not only effects against oxidative stress, but also various anti-inflammatory and anti-allergic effects. H2 regulates various gene expressions and protein-phosphorylations, though the molecular mechanisms underlying the marked effects of very small amounts of H2 remain elusive

The 2011 Medical Molecular Hydrogen Symposium: An inaugural symposium of the journal Medical Gas Research

This report summarizes a brief description/history of the Hydrogen Research Meetings as well as key presentations/oral abstracts delivered in the most recent symposium. Additionally, we introduced 38 diseases and physiological states for which hydrogen exhibits beneficial effects

Molecular hydrogen protects chondrocytes from oxidative stress and indirectly alters gene expressions through reducing peroxynitrite derived from nitric oxide

<p>Abstract</p> <p>Background</p> <p>Molecular hydrogen (H₂) functions as an extensive protector against oxidative stress, inflammation and allergic reaction in various biological models and clinical tests; however, its essential mechanisms remain unknown. H₂directly reacts with the strong reactive nitrogen species peroxynitrite (ONOO^-) as well as hydroxyl radicals (•OH), but not with nitric oxide radical (NO•). We hypothesized that one of the H₂functions is caused by reducing cellular ONOO^-, which is generated by the rapid reaction of NO• with superoxides (•O₂^-). To verify this hypothesis, we examined whether H₂could restore cytotoxicity and transcriptional alterations induced by ONOO^-derived from NO• in chondrocytes.</p> <p>Methods</p> <p>We treated cultured chondrocytes from porcine hindlimb cartilage or from rat meniscus fibrecartilage with a donor of NO•, <it>S</it>-nitroso-<it>N</it>-acetylpenicillamine (SNAP) in the presence or absence of H₂. Chondrocyte viability was determined using a LIVE/DEAD Viability/Cytotoxicity Kit. Gene expressions of the matrix proteins of cartilage and the matrix metalloproteinases were analyzed by reverse transcriptase-coupled real-time PCR method.</p> <p>Results</p> <p>SNAP treatment increased the levels of nitrated proteins. H₂decreased the levels of the nitrated proteins, and suppressed chondrocyte death. It is known that the matrix proteins of cartilage (including aggrecan and type II collagen) and matrix metalloproteinases (such as MMP3 and MMP13) are down- and up-regulated by ONOO^-, respectively. H₂restoratively increased the gene expressions of aggrecan and type II collagen in the presence of H₂. Conversely, the gene expressions of MMP3 and MMP13 were restoratively down-regulated with H₂. Thus, H₂acted to restore transcriptional alterations induced by ONOO^-.</p> <p>Conclusions</p> <p>These results imply that one of the functions of H₂exhibits cytoprotective effects and transcriptional alterations through reducing ONOO^-. Moreover, novel pharmacological strategies aimed at selective removal of ONOO^-may represent a powerful method for preventive and therapeutic use of H₂for joint diseases.</p

A novel high-throughput (HTP) cloning strategy for site-directed designed chimeragenesis and mutation using the Gateway cloning system

There is an increasing demand for easy, high-throughput (HTP) methods for protein engineering to support advances in the development of structural biology, bioinformatics and drug design. Here, we describe an N- and C-terminal cloning method utilizing Gateway cloning technology that we have adopted for chimeric and mutant genes production as well as domain shuffling. This method involves only three steps: PCR, in vitro recombination and transformation. All three processes consist of simple handling, mixing and incubation steps. We have characterized this novel HTP method on 96 targets with >90% success. Here, we also discuss an N- and C-terminal cloning method for domain shuffling and a combination of mutation and chimeragenesis with two types of plasmid vectors

両側Fan-shaped flap法で再建した下口唇有棘細胞癌の1例 : Fan-shaped flap作製上の工夫について

口唇癌の95%以上が下口唇に認められ,組織学的に90%以上が有棘細胞癌であるといわれている.外科的治療が下口唇癌に対ししばしば選択されるが,整容的な問題だけでなく食事・会話等の機能的問題も生じうる.下口唇組織は幅1/3までは単純縫縮が可能であるが欠損幅が1/3を超えると何らかの再建術が必要となる.開口機能,閉口機能,構音機能,感情表現,知覚を最大限再現することが術後機能回復の上で重要となる.皮膚・筋肉・粘膜の三層構造を持つ頬部や口唇の組織を全層で用いる局所皮弁は,機能的にも整容的にも良い結果を得やすい.今回われわれは下口唇に生じた有棘細胞癌に対し下口唇切除後,両側Fan-shaped flap法に工夫を加えて再建した.術後食事,会話に問題なく整容的にも満足する結果が得られた.It is known that &#8805; 95% of lip cancers occur on the lower lip, and that &#8805; 90% are histologically considered to represent squamous cell carcinoma. Surgical procedures are often selected for the treatment of lower lip cancer, but may result in not only cosmetic, but also functional issues, such as problems with meal intake and conversation. Simple closures up to one-third of the width can be used on lower lip tissue, but if the defect width exceeds this, some type of reconstruction becomes necessary. Maximal reproducibility of opening function, closing function, articulation function, emotional expression, and perception is essential for postoperative functional recovery. The local flap technique using all layers of the buccal and lip tissues, which have a three-layered structure consisting of skin, muscle, and mucosa, is known to offer greater success in achieving satisfactory functional and cosmetic results. Here, we describe lower lip resection and subsequent reconstruction using a bilateral fan-shaped flap technique with a modified approach in a patient with squamous cell carcinoma on the lower lip. No issues with meal intake or conversations were seen postoperatively, and satisfactory cosmesis was also achieved