[Minimally Invasive Aortic Valve Replacement for Aortic Valve Infective Endocarditis Complicated by Septic Arthritis of the Sternoclavicular Joint:Report of a Case].

A 54-year-old man with a history of atopic dermatitis was admitted to our hospital for persistent fever and multiple arthralgias unresponsive to antibiotics. On the second day of hospitalization, Staphylococcus aureus was detected in the blood culture, and debridement for presumed pyogenic arthritis was performed on the patient's bilateral wrists and right ankle joints. Echocardiography showed evidence of infective endocarditis of the aortic valve. The patient's fever persisted after drainage of multiple joint abscesses, and blood cultures remained positive. A right sternoclavicular joint abscess that had been noted on computed tomography (CT) at the time of admission had not decreased in size on repeat CT performed 10 days post-admission. After additional drainage of the sternoclavicular joint abscess on the 15th day, the patient's fever subsided, and blood culture was negative. On the 29th day, an aortic valve replacement was performed via a right anterior thoracotomy to prevent sternal osteomyelitis. The postoperative course was uneventful, and the patient was discharged on the 35th day after valve surgery. One year after the surgery, he continues to take antibiotics, and recurrence of infection has not been observed.

[Determination of Endograft Size in Thoracic Endovascular Aortic Repair for Blunt Thoracic Aortic Injury:Report of a Case].

Although the efficacy of thoracic endovascular aortic repair( TEVAR) for blunt thoracic aortic injury (BTAI) has been reported, the determination of the proper endograft size for TEVAR in BTAI cases is yet to be established. The management of BTAI by TEVAR may not be effective if a wrong endograft size is selected, possibly leading to fatal outcomes. In this case report, a hemodynamically unstable patient with BTAI was saved by TEVAR, and the preoperative and postoperative changes in the aortic diameter were compared. In TEVAR for BTAI, the effects of hemodynamic instability should be considered and the appropriate device size should be determined based on blood pressure, pulse, and inferior vena cava diameter.

[Clinical Features and Treatment Strategy of Vertebral Artery Injury Associated with Cervical Spine Trauma].

OBJECTIVE: Vertebral artery injury(VAI)associated with cervical spine trauma has the potential to cause catastrophic vertebrobasilar stroke. However, there are no well-defined treatment recommendations for VAI. The purpose of this study was to identify an effective treatment strategy for VAI following cervical spine trauma. METHODS: Ninety-seven patients with blunt cervical spine trauma were treated at Hyogo Prefectural Kakogawa Medical Center between January 2013 and September 2017. Of these patients, 49 underwent computed tomographic angiography or magnetic resonance angiography for evaluation of the vertebral artery. Eighteen patients(36.7%)had a diagnosis of VAI. We retrospectively analyzed the clinical features, treatment, and outcomes in these 18 patients. RESULTS: Seven patients(38.9%)had bilateral VAI, 16(88.9%)had cervical dislocation, and 2(11.1%)had transverse process fractures extending into the transverse foramen. Surgical reduction was performed in 14 patients. Five patients with either bilateral or unilateral occlusion underwent parent artery occlusion before reduction. There were no complications after this procedure. Two patients with bilateral VAI had a stroke before treatment. There were no infarctions in the distribution of the vertebrobasilar artery after intervention. The perioperative stroke rate was relatively good, and almost all Glasgow Outcome Scale scores were related to the degree of spinal cord injury. CONCLUSIONS: Aggressive screening for VAI is important in patients with cervical spine trauma in order to ensure adequate treatment. Although the treatment strategy described here could yield good results, it may require modification according to the needs of the individual patient.

[Endovascular Repair for Acute Phase of Retrograde Type A Aortic Dissection with an Entry in the Descending Aorta].

OBJECTIVE: Endovascular repair for retrograde type A aortic dissection with an entry in the descending aorta (RAAD) is challenging. We present early and mid- term results of endovascular repair for acute phase of RAAD by using commercially-available device. METHODS: From April 2012 to June 2014, 10 consecutive patients with acute phase of RAAD underwent endovascular repair in our hospital. Of them, 9 patients had emergency surgery within 24 hours after the onset. The other one patient had urgent surgery 3 days after the onset. In all patients, the entry tear was covered with TAG or conformable TAG. RESULTS: Technical success was achieved in all patients. No in-hospital mortality was experienced. In all patients, follow-up computed tomography images showed significant remodeling in the ascending aorta 3 months after surgery. During a median follow-up period of 19.5 months, no patients died and no re-intervention occurred. CONCLUSIONS: In patients with acute phase of RAAD, endovascular repair with commercially-available device can be safely performed and it provides sufficient remodeling in the ascending aorta early after surgery. This technique is an alternative to open repair in these patients.

[Adult endocardial blood cyst; report of a case].

We report herein a rare case of endocardial blood cyst (EBC) in an adult patient. A 63-year-old asymptomatic woman underwent echocardiography, which incidentally detected a cardiac tumor in the right atrium. On echocardiography, the tumor was revealed to be a 30-mm round mass with thin, hyperechogenic walls and heterogeneously hypoechogenic contents. The lesion was attached to the septum. On computed tomography, the tumor appeared partly calcified and showed poor contrast-enhancement. On magnetic resonance imaging, the lesion appeared isointense or slightly hyperintense in T1 and T2-weighted sequences. Myxoma was strongly suspected based on these preoperative imaging findings. The tumor was successfully excised under cardiopulmonary bypass. Gross examination confirmed that the cyst was filled with blood. The cystic walls comprised thin-layered fibrous tissue lined with endocardial cells. No tumor cells were found. The diagnosis of EBC was confirmed based on histopathological examination, and the postoperative course was uneventful.

Aspartic protease from Aspergillus (Eurotium) repens strain MK82 is involved in the hydrolysis and decolourisation of dried bonito (Katsuobushi).

BACKGROUND: Katsuobushi is a dried, smoked and fermented bonito used in Japanese cuisine. During the fermentation process with several Aspergillus species, the colour of Katsuobushi gradually changes from a dark reddish-brown derived from haem proteins to pale pink. The change in colour gives Katsuobushi a higher ranking and price. This study aimed to elucidate the mechanism of decolourisation of Katsuobushi. RESULTS: A decolourising factor from the culture supernatant of Aspergillus (Eurotium) repens strain MK82 was purified to homogeneity. The purification was monitored by measuring the decolourising activity using equine myoglobin and bovine haemoglobin as substrates. It was found that the decolourising factor had protease activity towards myoglobin and haemoglobin. Complete inhibition of the enzyme by the inhibitor pepstatin A and the internal amino acid sequence classified the protein as an aspartic protease. The enzyme limitedly hydrolysed myoglobin between 1-Met and 2-Gly, 43-Lys and 44-Phe, and 70-Leu and 71-Thr. The purified enzyme decolourised blood of Katsuwonus pelamis (bonito) and a slice of dried bonito. CONCLUSION: It is proposed that aspartic protease plays a role in the decolourisation of Katsuobushi by the hydrolysis of haem proteins that allows the released haem to aggregate in the dried bonito.

Takotsubo cardiomyopathy associated with hypoglycemia: inverted takotsubo contractile pattern.

Classic takotsubo cardiomyopathy (TCM) is characterized by transient dysfunction of the apical portion of the left ventricle with hyperkinesis of the other parts of the heart wall. Recently, wall motion abnormalities in parts other than in the apical portion of the heart have been reported. Inverted TCM is one form of these anomalies. In this form, the basal segments rather than the apical part of the heart are akinetic/dyskinetic, and the apex is hyperdynamic. Emotional or physical stress is known to trigger TCM, leading some authors to call TCM stress-induced cardiomyopathy (SC). Hypoglycemia is regarded as one of the physical stresses that cause TCM/SC. We present a case of inverted TCM/SC with hypoglycemia. In this case, a 60-year-old woman was brought to our hospital with loss of consciousness caused by hypoglycemia. Initially, the echocardiography revealed an inverted takotsubo contractile pattern. The patient was stabilized with continuous intravenous fluids and a glucose injection, whereas the echocardiography on day 4 showed an almost normal contractile pattern. Among the case reports regarding hypoglycemia as a preceding stressor of TCM/SC, a case of inverted TCM/SC with hypoglycemia is rare. Hypoglycemia is a relatively common case in emergency department; however wall motion abnormalities are not usually expected in hypoglycemic patients. Thus, undiagnosed self-limited TCM/SC cases are possible among hypoglycemic patients. TCM/SC is reported to be a cause of torsade de pointes, which can be fatal. This might warrant an echocardiogram for hypoglycemic patients so as not to overlook TCM/SC in the emergency department.

Organic solvent-tolerant elastase efficiently hydrolyzes insoluble, cross-linked, protein fiber of eggshell membranes.

Eggshell membrane is a mechanically stable and insoluble cross-linked fibrous protein. Pseudomonas aeruginosa strain ME-4 synthesizes a metalloprotease that degrades the eggshell membrane. We cloned the encoding gene in Escherichia coli. The recombinant protease, over-expressed in E. coli, was inactive but addition of acetone to crude cell extracts restored the activity and removed many E. coli proteins. We purified the active, acetone-treated protease to homogeneity in a single chromatography step with 57% recovery. The recombinant protease partially hydrolyzed eggshell membrane and produced more soluble peptides and proteins than commercial elastase, α-chymotrypsin, and collagenase. The soluble peptides produced from hydrolyzed eggshell membrane inhibited angiotensin-I-converting enzyme activity. The degradation of eggshell membrane by the recombinant elastase could be applied to the production of soluble bioactive peptides.

Ampicillin/sulbactam versus cefazolin plus aminoglycosides for antimicrobial prophylaxis in management of Gustilo type IIIA open fractures: A retrospective cohort study.

BACKGROUND: The antibiotic regimens for prophylaxis in the management of open fractures remain controversial. Although the use of aminoglycosides is widely accepted for treatment of Gustilo type III open fractures, aminoglycosides are often avoided in patients with risk factors. This study aimed to compare efficacy and safety of two regimens, cephazolin plus aminoglycoside (amikacin or gentamicin) and ampicillin/sulbactam (ABPC/SBT), in patients with Gustilo type IIIA open fractures. METHODS: A total of 95 Gustilo type IIIA fractures in 90 patients were retrospectively reviewed in this study. The cohort was categorized into two groups that were treated in accordance with the institutional prescribed regimen in different periods: (1) cefazolin plus aminoglycoside (January 1, 2014-September 30, 2017) and (2) ABPC/SBT monotherapy (October 1, 2017-September 30, 2020). Cefazolin was used at 1-2 g every 8 h, aminoglycoside (amikacin or gentamicin) was used daily depending on body weight, and ABPC/SBT was used at 3 g every 8 h The antibiotic administration was continued within 3 days or until successful soft tissue coverage was achieved. The infection rate and the incidence of acute kidney injury (AKI) in both groups were assessed. RESULTS: ABPC/SBT was used in 34 patients (36 fractures), and 56 patients (59 fractures) received cefazolin plus aminoglycoside for antibiotic prophylaxis. Infection developed in 2 of 36 fractures in ABPC/SBT group and 4 of 59 fractures in the cefazolin plus aminoglycoside group (p > 0.99). The average serum creatinine levels on admission, baseline, and peak during the hospital stay were not significantly different between the two groups. One case of AKI was identified in each group, indicating that incidence rate of AKI was not significantly different between the two groups. CONCLUSION: We demonstrated the non-inferiority of ABPC/SBT therapy over cefazolin plus aminoglycoside regimen for type IIIA open fractures. The ABPC/SBT regimen may be an alternative option for managing Gustilo type IIIA open fractures. Further prospective studies with larger samples are needed to verify these results.

Characterization of YIPF3 and YIPF4, cis-Golgi Localizing Yip domain family proteins.

The Yip1 domain family (YIPF) proteins are homologues of yeast Yip1p and Yif1p, which are proposed to function in ER to Golgi transport. Here, we report the characterization of YIPF3 and YIPF4, homologues of human Yif1p and Yip1p, respectively. Immunofluorescence and immuno-electron microscopy showed that both YIPF3 and YIPF4 are clearly concentrated in the cis-Golgi. While YIPF4 was detected as a single mobility form consistent with its predicted molecular weight, three different mobility forms of YIPF3 were detected by western blotting. Biochemical and immunofluorescence experiments strongly indicated that YIPF3 is synthesized in the ER as a N-glycosylated form (40 kDa), is then O-glycosylated in the Golgi apparatus to become a lower mobility form (46 kDa) and finally becomes a higher mobility form cleaved at its C-terminal luminal domain (36 kDa). YIPF3 and YIPF4 form a complex in the Golgi apparatus, and this was suggested to be important for their proper localization and function. The knockdown of YIPF3 or YIPF4 in HeLa cells induced fragmentation of the Golgi apparatus, suggesting their involvement in the maintenance of the Golgi structure.

Molecular cloning and sequence analysis of two distinct halotolerant extracellular proteases from Bacillus subtilis FP-133.

We cloned the genes encoding the two distinct extracellular halotolerant proteases of Bacillus subtilis FP-133 Expro-I and Expro-II, which were classified as alkaline serine and neutral proteases respectively. Three-dimensional modeling suggested that acidic and polar amino acid residues located on the surface stabilize protein structure in the presence of relatively high NaCl concentrations.

Fe-superoxide dismutase and 2-hydroxy-1,4-benzoquinone reductase preclude the auto-oxidation step in 4-aminophenol metabolism by Burkholderia sp. strain AK-5.

Burkholderia sp. strain AK-5 converts 4-aminophenol to maleylacetic acid via 1,2,4-trihydroxybenzene, which is unstable in vitro and non-enzymatically auto-oxidized to 2-hydroxy-1,4-benzoquinone. Crude extract of strain AK-5 retarded the auto-oxidation and reduced the substrate analogue, 2,6-dimethoxy-1,4-benzoquinone, in the presence of NADH. The two enzymes responsible were purified to homogeneity. The deduced amino acid sequence of the enzyme that inhibited the auto-oxidation showed a high level of identity to sequences of iron-containing superoxide dismutases (Fe-SODs) and contained a conserved metal-ion-binding site; the purified enzyme showed superoxide dismutase activity and contained 1 mol of Fe per mol of enzyme, identifying it as Fe-SOD. Among three type SODs tested, Fe-SOD purified here inhibited the auto-oxidation most efficiently. The other purified enzyme showed a broad substrate specificity toward benzoquinones, including 2-hydroxy-1,4-benzoquinone, converting them to the corresponding 1,4-benzenediols; the enzyme was identified as 2-hydroxy-1,4-benzoquinone reductase. The deduced amino acid sequence did not show a high level of identity to that of benzoquinone reductases from bacteria and fungi that degrade chlorinated phenols or nitrophenols. The indirect role of Fe-SOD in 1,2,4-trihydroxybenzene metabolism is probably to scavenge and detoxify reactive species that promote the auto-oxidation of 1,2,4-trihydroxybenzene in vivo. The direct role of benzoquinone reductase would be to convert the auto-oxidation product back to 1,2,4-trihydroxybenzene. These two enzymes together with 1,2,4-trihydroxybenzene 1,2-dioxygenase convert 1,2,4-trihydroxybenzene to maleylacetic acid.

Expression and characterization of a fusion protein-containing cyclodextrin glycosyltransferase from Paenibacillus sp. A11.

A recombinant cyclodextrin glycosyltransferase (CGTase) gene fused with thioredoxin (Trx), hexa-histidine (His(6)) and S-protein (S) at the N terminus and a proline-rich peptide (PRP) at the C terminus, was constructed using the wild-type gene from Paenibacillus sp. A11, the pET-32a vector and Escherichia coli BL21(DE3) as the host cell. The expression levels and enzyme characteristics of the Trx-His(6)-CGTase-PRP fusion protein, the recombinant CGTase without fusion peptides, and the wild-type CGTase were compared. The maximum specific activity for the Trx-His(6)-CGTase-PRP fusion enzyme was 2.7 fold higher than that of the non-fusion form at the optimal IPTG concentration. The Trx-His(6)-CGTase-PRP fusion protein was purified to homogeneity by starch adsorption and Ni-NTA affinity chromatography, with a specific activity of 2,268 units/mg protein at a 61% yield. The ease of purification and the higher enzyme yield were obtained with the fusion form when compared to the non-fusion and wild-type enzymes. The fusion enzyme was superior than its wild-type counterpart in terms of stability against high temperature and organic solvents. Moreover, the fusion enzyme could catalyze the synthesis of cyclodextrins in 20% (v/v) dimethylformamide with a higher product yield of CD(7) and CD(8) compared to that of the wild-type enzyme in the same buffer-solvent system.

[Efficacy of vacuum-assisted closure therapy for various non-healing wounds after cardiovascular and thoracic surgery].

Vacuum-assisted closure (VAC) therapy is an efficacious modality for treating chronic and difficult wounds. We present 3 cases that responded well to VAC therapy after cardiovascular and thoracic surgery: 1 methicillin-resistant Staphylococcus aureus (MRSA) wound infection after Stony's incision, 1 inguinal lymphorrhea, and 1 empyema after a traffic accident The duration of VAC therapy was 9, 18, and 90 days, respectively, and all 3 wounds healed completely. Familiar equipment and supplies available on the hospital ward were used, and patients were able to leave their beds. In this report, the efficacy of VAC therapy, the problems encountered, and the steps that could be taken to address them are discussed.

Purification and characterization of an extracellular laccase from Phlebia radiata strain BP-11-2 that decolorizes fungal melanin.

Phlebia radiata strain BP-12-2, isolated from forest soil, decolorized fungal melanin and produced an extracellular laccase in culture. The enzyme was purified to homogeneity and characterized. It showed no absorption bands in the ultraviolet above 300 nm or visible regions. It differed from the reported laccases in substrate specificity, kinetic parameters, and NH2-terminal amino acid sequences.

Purification, characterization, and gene cloning of Ceriporiopsis sp. strain MD-1 peroxidases that decolorize human hair melanin.

Ceriporiopsis sp. strain MD-1, isolated from forest soil, produced several extracellular enzymes that decolorized human hair melanin. Among them, three enzymes (E1, E2-1, and E2-2) were purified to homogeneity and characterized. The enzymes required hydrogen peroxide in their enzyme reactions and, typical of other fungal peroxidases, oxidized various phenol compounds such as guaiacol, but not 3,4-dimethoxybenzyl alcohol. The spectra of the three enzymes showed an absorption maximum at 406 nm, indicating that they were heme proteins. However, the A(406)/A(280) values of the enzymes were below 0.4, which was lower than those of other peroxidases. E2-1 and E2-2 were similar to each other in their molecular and catalytic properties, and they possibly represent products of posttranslational modifications and/or allelic variants of the same gene, mdcA. The corresponding cDNA was cloned and sequenced; the deduced amino acid sequence showed high identities to the manganese peroxidases from other microorganisms. The specific activities and K(m) values of E2-1 and E2-2 for synthetic and human hair melanins were much higher than those of Phanerochaete chrysosporium manganese peroxidase and lignin peroxidase.

Cloning and expression of cyclodextrin glycosyltransferase gene from Paenibacillus sp. T16 isolated from hot spring soil in northern Thailand.

Gene encoding cyclodextrin glycosyltransferase (CGTase), from thermotolerant Paenibacillus sp. T16 isolated from hot spring area in northern Thailand, was cloned and expressed in E. coli (JM109). The nucleotide sequences of both wild type and transformed CGTases consisted of 2139 bp open reading frame, 713 deduced amino acids residues with difference of 4 amino acid residues. The recombinant cells required 24 h culture time and a neutral pH for culture medium to produce compatible amount of CGTase compared to 72 h culture time and pH 10 for wild type. The recombinant and wild-type CGTases were purified by starch adsorption and phenyl sepharose column chromatography and characterized in parallel. Both enzymes showed molecular weight of 77 kDa and similar optimum pHs and temperatures with recombinant enzyme showing broader range. There were some significant difference in pH, temperature stability and kinetic parameters. The presence of high starch concentration resulted in higher thermostability in recombinant enzyme than the wild type. The recombinant enzyme was more stable at higher temperature and lower pH, with lower K(m) for coupling reaction using cellobiose and cyclodextrins as substrates.

Metabolism of azo dyes by Lactobacillus casei TISTR 1500 and effects of various factors on decolorization.

Lactobacillus casei TISTR 1500 was isolated from soil of a dairy wastewater treatment plant and selected as the most active azo dye degrader of 19 isolates. Growing cells and freely suspended cells of this strain completely degraded methyl orange, thereby decolorizing the medium. The strain stoichiometrically converted methyl orange to N,N-dimethyl-p-phenylenediamine and 4-aminobenzenesulfonic acid, which were identified by HPLC, GC, and GC-MS analyses. The enzyme activity responsible for the cleavage of the azo bond of methyl orange was localized to the cytoplasm of cells grown on modified MRS medium containing methyl orange. The effect of sugars, oligosaccharides, organic acids, metal ions, pHs, oxygen and temperatures on methyl orange decolorization by freely suspended cells was investigated. The optimal conditions for the decolorization of methyl orange by the Lactobacillus casei TISTR 1500 are incubation at 35 degrees C and pH 6 with sucrose provided as the energy source.