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Cadherin 23 (CDH23) is one of the most common genes responsible for hereditary hearing loss; a mutation of CDH23 can cause a wide range of symptoms depending on the variant. In this study, an iPSC line was generated from a patient with late-onset, progressive high frequency hearing loss caused by c.[719C > T];[6085C > T]:p.[P240L];[R2029W] compound heterozygous variants of CDH23. The cells were confirmed to have a normal karyotype, express markers of pluripotency, and have tri-embryonic differentiation potential. This disease-specific iPSC line will further the construction of disease models and the elucidation of the pathophysiology of CDH23 mutations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Respiratory syncytial virus (RSV) and rotavirus infections are long-standing infectious diseases that affect children worldwide. RSV and rotavirus were first discovered in clinical specimens in 1955 and 1973, respectively. From their discovery to the present day, significant progress has been made in understanding these two infections. The introduction of a simple and rapid antigen diagnostic test into clinical settings in the 1990s offered new insight into the clinical characteristics and epidemiology of these infections. Regarding therapeutics, symptomatic treatments have remained the mainstay; however, prophylactic humanized anti-RSV monoclonal antibodies have been developed and advances in structural biology may allow for more effective human anti-RSV monoclonal antibodies and novel RSV vaccines to be developed soon. For rotavirus, two vaccines have been licensed and broadly applied over the past 10 years, which have been successful clinically and have changed the epidemiology of rotavirus infections in Japan., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2024 Japanese Society of Chemotherapy, Japanese Association for Infectious Diseases, and Japanese Society for Infection Prevention and Control. Published by Elsevier Ltd. All rights reserved.)

Usher syndrome type 2A (USH2A) gene mutations have been identified as the most frequent genetic causes of hereditary deafness in Usher syndrome, and an effective treatment has yet to be established. The encoded protein, Usherin, is essential for the ankle link associated with extracellular connections between the stereocilia of inner ear hair cells. We report the generation of a patient-derived USH2A iPSC line with compound mutations c.1907_1912ATGTTT > TCACAG (p.D636V + V637T + C638G) and c.8328_8329delAA (p.L2276fs*12). The iPSC showed the expression of pluripotency markers, the ability to differentiate into three germ layers in vitro, and USH2A mutations with normal karyotype., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

The increasing eutrophication of freshwater and brackish habitats globally has led to a corresponding increase in the occurrence of harmful cyanobacterial blooms. Cyanobacteria can produce highly toxic substances such as microcystins (MCs) that affect the health of livestock, wildlife, and humans. The present study broaden the understanding of cyanobacteria ecology and MC dynamics in the field, focusing on the estimation of the production and sedimentation rates of MCs in a natural habitat. The nutrient concentrations of the reservoir water and sediment pore water were monitored at 3-h intervals for 24 h during the summer cyanobacterial bloom. The DIN uptake rate of Microcystis in the Isahaya reservoir was estimated and the large-scale blooms in the reservoir were largely controlled by the interactions between rainfall and nutrient levels in the warm season. By using calculations based on the nitrogen budgets and tracking changes of the MC concentrations in the water column, the total MC production and sedimentation rates were estimated to be 52.2 kg MCs d -1 and 21.5 kg MCs d -1 , respectively. Although MCs could be degraded in the environment, the MC sedimentation still comprised 41% of the in-water production., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Half a century has passed since the department for education and research on forensic odontology was established at dentistry-related universities in Japan in 1964. In order to meet the demands of society, the number of universities with a department of forensic odontology increased up until around 2005. In 2007, the Japanese Society of Forensic Dental Science was established, and then a series of reforms such as establishment of the Study Council on Death Cause Investigation in both the National Police Agency and the Cabinet Office of the Japanese government, cabinet decision of enactment and enforcement of new laws on death cause investigation, publication of an article on the Model Core Curriculum of Dental Education, publication of the results of a fact-finding survey on education and research on forensic odontology conducted by the Ministry of Education, Culture, Sports, Science and Technology, inclusion of questions about forensic odontology in the National Board Dental Examination, and compilation of a database on dental findings by the Ministry of Health, Labor and Welfare, proceeded in succession. We introduced the half century of forensic odontology in Japan in chronological order., (© 2019 The Authors.)

A 13-year-old boy developed tetanus, although he had protective antitoxin antibody raised by three doses of tetanus toxoid vaccine. Four days after injury, he presented with muscle rigidity of his posterior neck, excessive diaphoresis, and risus sardonicus and was subsequently diagnosed with tetanus. Tetanus is rare in developed countries, particularly during childhood, but must be promptly diagnosed based on clinical symptoms., (Copyright © 2018 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

Aims: We conducted a longitudinal cohort study to analyze the relationship between outcome of gross motor development in preterm infants and factors that might affect their development., Methods: Preterm infants with a birth weight of <1500 g were recruited. We measured spontaneous antigravity limbs movements by 3D motion capture system at 3 months corrected age. Gross motor developmental outcomes at 6 and 12 months corrected age were evaluated using the Alberta Infant Motor Scale (AIMS). Statistical analysis was carried out by canonical correlation analysis., Results: Eighteen preterm infants were included. In the 6 months corrected age analysis, spontaneous movement had a major effect on Prone and Sitting at 6 months corrected age of AIMS. In the 12 months corrected age analysis, spontaneous movement had a major effect on Sitting and Standing at 12 months corrected age of AIMS., Conclusions: In preterm infants, better antigravity spontaneous movements at 3 months corrected age were significantly correlated with better gross motor development at 6 or 12 months corrected age., (Copyright © 2018 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

Background: Periostin and squamous cell carcinoma antigen (SCCA) are involved in the pathogenesis of asthma. Acute bronchitis due to respiratory syncytial virus (RSV) infection during infancy exhibits an asthma-like pathogenesis, suggesting that it may be associated with the subsequent development of asthma. However, the mechanism by which RSV infection leads to development of asthma has not yet been fully elucidated., Methods: Infants younger than 36 months were enrolled and classified into three groups. Group I included patients hospitalized with RSV-induced bronchitis. These patients were further stratified into two sub-groups according to whether the criteria for the modified Asthma Predictive Index (mAPI) had been met: Group I consisted of mAPI (+) and mAPI (-) patients; Group II included patients with food allergy as a positive control group; and Group III included children with no allergy as a negative control group. Serum periostin and SCCA levels were measured in the groups. This study was registered as a clinical trial (UMIN000012339)., Results: We enrolled 14 subjects in Group I mAPI (+), 22 in Group I mAPI (-), 18 in Group II, and 18 in Group III. In Group I, the serum periostin and SCCA levels were significantly higher during the acute phase compared with the recovery phase. However, no significant differences were found between Group I mAPI (+) and mAPI (-)., Conclusions: The serum periostin and SCCA levels increased during acute RSV bronchitis. Both periostin and SCCA may play a role in the pathogenesis of acute bronchitis due to RSV., (Copyright © 2018 Japanese Society of Allergology. Production and hosting by Elsevier B.V. All rights reserved.)

ST121/ agr -IV methicillin-susceptible Staphylococcus aureus was isolated from a patient of septic arthritis (synovial fluid, blood, skin and nasal cavity). Although the Panton-Valentine leukocidin (PVL) gene was negative, this isolate harboured a gene encoding a variant of bone sialoprotein-binding protein with a shortened SD-repeat region.

Macrolide antibiotics have immunomodulatory activities, including suppression of cytokine production, cell adhesion molecule expression, and mucin production. These immunomodulatory activities improve the symptoms of respiratory diseases associated with chronic inflammation. However, the underlying molecular mechanism(s) is not well understood yet. To address this, we prepared clarithromycin (CAM)-conjugated Sepharose and examined bound cellular proteins by proteome analysis. We identified mitochondrial proteins 4-nitrophenylphosphatase domain and non-neuronal synaptosomal associated protein 25-like protein homolog (NIP-SNAP)-1 and -2 and very long-chain acyl-CoA dehydrogenase (VLCAD) as CAM-binding proteins. Production of proinflammatory cytokines (IL-8 and IL-6) induced by lipopolysaccharides (LPSs) and Pam 3 -CSK 4 in human epithelial cell lines BEAS-2B and T24 were suppressed by knockdown of NIP-SNAP-1 or -2, and partly by knockdown of VLCAD. Also, knockdown of NIP-SNAP-1 or -2 in various cell lines suppressed LPS-induced expression of IL-8 and IL-6 mRNA and NF-κB activity. Thus, CAM suppresses NF-κB-mediated proinflammatory cytokine production by interacting with mitochondrial proteins, NIP-SNAP-1 and -2., (Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.)

NIP-SNAP-1 and -2 are ubiquitous proteins thought to be associated with maintenance of mitochondrial function, neuronal transmission, and autophagy. However, their physiological functions remain largely unknown. To elucidate their functional importance, we screened for proteins that interact with NIP-SNAP-1 and -2, resulting in identification of HSP60 and P62/SQSTM1 as binding proteins. NIP-SNAP-1 and -2 localized in the mitochondrial inner membrane space, whereas HSP60 localized in the matrix. Native gel electrophoresis and filter trap assays revealed that human HSP60 prevented aggregation of newly synthesized NIP-SNAP-2 in an in vitro translation system. Moreover, expression levels of NIP-SNAP-1 and -2 in cells were decreased by knockdown of HSP60, but not HSP10. These findings indicate that HSP60 promotes folding and maintains the stability of NIP-SNAP-1 and -2., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Macrolide antibiotics exert immunomodulatory activity by reducing pro-inflammatory cytokine production by airway epithelial cells, fibroblasts, vascular endothelial cells, and immune cells. However, the underlying mechanism of action remains unclear. Here, we examined the effect of clarithromycin (CAM) on pro-inflammatory cytokine production, including interferons (IFNs), by primary human nasal epithelial cells and lung epithelial cell lines (A549 and BEAS-2B cells) after stimulation by Toll-like receptor (TLR) and RIG-I-like receptor (RLR) agonists and after infection by human respiratory syncytial virus (RSV). CAM treatment led to a significant reduction in poly I:C- and RSV-mediated IL-8, CCL5, IFN-β and -λ production. Furthermore, IFN-β promoter activity (activated by poly I:C and RSV infection) was significantly reduced after treatment with CAM. CAM also inhibited IRF-3 dimerization and subsequent translocation to the nucleus. We conclude that CAM acts a crucial modulator of the innate immune response, particularly IFN production, by modulating IRF-3 dimerization and subsequent translocation to the nucleus of airway epithelial cells. This newly identified immunomodulatory action of CAM will facilitate the discovery of new macrolides with an anti-inflammatory role., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A methicillin-susceptible Staphylococcus aureus with Panton-Valentine leukocidin (PVL) genes was isolated from refractory breast abscesses of 12-year-old girl in Japan, and classified into ST88, spa-t1245 and coa-IIIa. This strain harboured PVL phage ΦSa2usa, which is usually found in ST8 community-acquired methicillin-resistant S. aureus clone USA300.

Freshwater cyanobacteria produce highly toxic substances such as microcystins (MCs), and water containing MCs is often discharged to downstream and coastal areas. We conducted field monitoring in Isahaya Bay to clarify the short-term dynamics of MCs discharged from a reservoir following a cyanobacteria bloom in the warm season. MCs were detected in the seawater of the bay (max. 0.10 μg L(-1)), and were deposited on the sea floor, with the MC content of the surface sediment increasing by approximately five times (0.11±0.077-0.53±0.15 μg kgww(-1), mean±SD) at the four stations near the reservoir drainage gate before and after the discharge. The MCs was then transported from the mouth of the bay by tidal currents during the period of the study. Therefore, the MCs were moved away from the closed water area where the cyanobacteria blooms, and spread throughout the coastal area., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In the artificial reservoir of the Isahaya reclaimed land, Nagasaki, Japan, algal blooms have become an annual event, dominated primarily by the microcystin (MC) producing cyanobacteria Microcystis aeruginosa. Although the majority of MCs are either degraded by bacteria or washed out to sea, some remain in the sediment of the reservoir and bay throughout the year. As a result, they also accumulate in aquatic organisms (mullet, oyster, etc.) that inhabit the reservoir and surrounding areas, as well as midge flies that spend their larval period in the bottom of the reservoir. Accordingly, MCs also accumulate in the predators of these organisms, allowing the toxin to spread from the hydrosphere to terrestrial ecosystems. The most effective method for resolving this potentially dangerous condition is to introduce seawater into the reservoir by opening the drainage gates at high tide., (Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.)

A new method to evaluate endothelial function, namely, reactive hyperemia peripheral arterial tonometry (RH-PAT), has been developed. RH-PAT is an index of endothelial function, indicating initial atherosclerotic lesions. The present study aimed to investigate the effect of lifestyle modification with a focus on exercise training on RH-PAT in obese patients. We studied 43 obese patients (body mass index ≥ 30). RH-PAT was measured, and the RH-PAT index was calculated as a ratio of the digital pulse volume during reactive hyperemia divided by that at baseline. Further, we assessed body composition, arterial stiffness, insulin resistance, adipocytokine levels, and exercise tolerance. The exercise program consisted of 30 min on a cycle ergometer or treadmill, 3 times per week for 6 months. Training intensity was adjusted to the anaerobic threshold. Significant improvements were observed in the RH-PAT index following exercise training. We noted a significant reduction in weight, body fat percentage, and leptin values, and a significant increase in adiponectin levels and exercise tolerance. An abnormal baseline RH-PAT index was observed in 24 patients (55.8%); however, the improvement rate was higher in these patients than in patients with normal RH-PAT index values. Stepwise multiple regression analysis revealed that changes in insulin resistance (Δ”HOMA-IR) were independently correlated with changes in the RH-PAT index. Our results indicate that lifestyle modification with a focus on exercise training improved the RH-PAT index in obese patients. Patients with abnormal RH-PAT index values before lifestyle modification with exercise training demonstrated a high rate of improvement following exercise. Further, our results suggest that insulin resistance was the only independent factor influencing improvement in endothelial function., (© 2014 Asian Oceanian Association for the Study of Obesity . Published by Elsevier Ltd. All rights reserved.)

Pelizaeus-Merzbacher disease (PMD) is a rare dysmyelinating disorder caused by mutations in the proteolipid protein 1 (PLP1) gene. PMD is generally classified according to its clinical or pathological features into classical or connatal forms. We describe here a 19-year-old male with classical form PMD who presented with stridor and nystagmus in early infancy and whose psychomotor development has been severely delayed. Brain magnetic resonance imaging revealed white matter abnormalities typical of PMD. Direct sequencing of the PLP1 gene identified two nucleotide substitutions. One was a C-to-T transition at -31 in the 5'-flanking region of exon 1; the other was a novel point mutation, T-to-C transition in exon 4, which led to substitution of cysteine for arginine at residue 184. Because Cys184 forms a disulphide bridge with Cys228, the Cys184Arg mutation probably removes the bridge and changes the tertiary structure of PLP protein. A defective disulfide bond in PLP protein could be important in the pathogenesis of PMD., (Copyright © 2010 The Japanese Society of Child Neurology. Published by Elsevier B.V. All rights reserved.)

In Japan, palivizumab, a humanized monoclonal antibody specific for respiratory syncytial virus (RSV), has been available since 2002. However, its use is limited to children at risk of severe RSV infection, with specific criteria that have been validated in large-scale clinical studies. The Pharmaceutical Committee of the Japan Pediatric Society established a committee to conduct a nationwide questionnaire survey to determine which diseases place children at risk of severe RSV infection and require preventive measures. A questionnaire sent to 613 medical institutions, including major pediatric hospitals and general hospitals with pediatric services, received 272 responses (44.4%). In total, 1,115 children not meeting current indications for palivizumab therapy were hospitalized for severe RSV infection, 16 (1.4%) of whom died; this suggests that palivizumab therapy should be considered for children with severe immunodeficiency or those at risk of nosocomial RSV infection in whom prevention of RSV infection by standard control measures appears difficult.

Porous metallic materials can have a low Young's modulus, which is approximately equal to that of human bone, by controlling the porosity. On the other hand, certain medical polymers exhibit biofunctionalities that are not intrinsically present in metallic materials. Therefore, a composite consisting of these materials is expected to possess both these advantages for biomedical applications. However, in the case of using porous metallic materials, the deterioration of mechanical properties should of concern because a stress concentration may be induced near the pores. In this study, for the fabrication of the abovementioned composite, a versatile process for filling a medical polymer into a porous metallic material has been developed using porous pure titanium (pTi) and polymethylmethacrylate (PMMA). Then, the tensile strength and Young's modulus of pTi filled with PMMA (pTi/PMMA) fabricated using this process are systematically investigated. The tensile strength of pTi can be improved by the PMMA filling. Particularly, the improvement in the tensile strength of pTi pretreated using a silane coupling agent before PMMA filling is greater than that of the non-pretreated pTi because the stress concentration near the pores may be reduced by the improvement in the interfacial adhesiveness between the titanium particles and the PMMA. In contrast, the effect of the PMMA filling on the Young's modulus of pTi is smaller than that on the tensile strength because the Young's modulus of PMMA is considerably lower than that of pTi. Further, tensile strengths and Young's moduli comparable to the tensile strength and Young's modulus of the human bone are successfully obtained in the case of some pTi/PMMA samples.

Sex determination from dental pulp DNA was examined by loop-mediated isothermal amplification (LAMP) method. Amelogenin locus was analyzed for sex determination. A set of four specially designed primers was prepared based on database from Gene Bank, and loop primers were designed to shorten the analysis time. Analysis was performed using 32 dental pulp DNA samples removal from permanent teeth stored at room temperature for 1-25 years after extraction. The X allele was detected in approximately 32min with real-time turbidimeter and the Y allele was detected in approximately 34min. Analysis time was reduced to half when using loop primers. Visual detection was also possible as the amplified product showed white turbidity. Sex determination by LAMP method was rapid and simple, and it should prove useful in unknown bodies of mass disasters.

Although graft-versus-host disease (GVHD) is a life-threatening complication of hematopoietic stem-cell transplantation (HSCT), its current diagnosis depends mainly on clinical manifestations and invasive biopsies. Specific biomarkers for GVHD would facilitate early and accurate recognition of this grave condition. Using proteomics, we screened for plasma proteins specific for GVHD in a mouse model. One peak with 8972-Da molecular mass (m/z) retained a discriminatory value in 2 diagnostic groups (GVHD and normal controls) with increased expression in the disease and decreased expression during cyclosporin A treatment, and was barely detectable in syngeneic transplantation. Purification and mass analysis identified this molecule as CCL8, a member of a large chemokine family. In human samples, the serum concentration of CCL8 correlated closely with GVHD severity. All non-GVHD samples contained less than 48 pg/mL (mean +/- SE: 22.5 +/- 5.5 pg/mL, range: 12.6-48.0 pg/mL, n = 7). In sharp contrast, CCL8 was highly up-regulated in GVHD sera ranging from 52.0 to 333.6 pg/mL (mean +/- SE: 165.0 +/- 39.8 pg/mL, n = 7). Strikingly, 2 patients with severe fatal GVHD had extremely high levels of CCL8 (333.6 and 290.4 pg/mL. CCL8 is a promising specific serum marker for the early and accurate diagnosis of GVHD.

We monitored seasonal changes of the abundance and composition of microorganisms in the fish-farm sediment in Kusuura Bay, Amakusa, Japan, using the quinone profiling technique, during bioremediation by introducing cultured colonies of polychaete, Capitella sp. I. In November 2004, approximately 9.2 million cultured worms were transferred to the fish-farm sediment, which increased rapidly, and reached 458.5 gWW/m(2) (528,000 indiv./m(2)) in March 2005. During this fast-increasing period of Capitella, the microbial quinone content of the surface sediment (0-2 cm) also increased markedly, and reached 237 micromol/m(2) in January 2005, although the water temperature decreased to the lowest levels in the year. Particularly, the mole fraction of ubiquinone-10 in total quinones in the sediment, indicating the presence of alpha subclass of Proteobacteria, increased by 9.3%. These facts suggest that the bacterial growth was enhanced markedly by the biological activities of worms in the sediment, and the bacteria played an important role in the decomposition of the organic matter in the sediment.

A polychaete, Capitella sp. I has been shown to degrade organics actively in organically enriched sediment below fish farms. Our aim of the present study is to enhance the biological treatment of sediment by co-inoculation of Capitella sp. I with bacterial isolates that possess high degrading potential for organic matter. We isolated a total of 200 bacterial strains from fecal pellets, burrow lining, worm body, and sediment, and selected six of them for the degradation experiments in the sediment microcosms. With two out of the six isolates, tentatively identified as Vibrio sp. and Vibrio cyclitrophicus by 16SrDNA sequence, we found the TOC reduction rate was stimulated in sediment co-inoculated with the worms and each of the bacteria. In contrast, this was not observed in sediments inoculated only with the worms or the bacterium. These results strongly suggest that co-inoculation of Capitella sp. I with bacteria improves biodegradation.

We have developed a "microscopic bubble generating system for the fish farm" and evaluated its ability to improve the quality of seawater and costs of it in marine cages of red sea bream (Pagrus major) in Kusuura Bay, Japan. Our results revealed that DO concentration of bubbling net pens increased and became significantly higher than the level outside the net pen (between 0.52 and 0.87 mg/L), and the whole water column was nearly saturated. Temperatures of the bubbling net pens decreased slightly between 0.08 and 0.12 degrees C in all the layers. Furthermore, micro-bubbles seemed to reach the deeper water due to the downward flow and diffusion. This study demonstrated that the microscopic bubble generating system developed in our research project could increase efficiently the dissolved oxygen concentration throughout all water layers of the fish farm. A capital and operation costs of the system is recoverable within a year.

Background: We developed a non-invasive, continuous, high-resolution method of measuring carboxyhaemoglobin fraction (COHb%) using expiratory gas analysis (EGA). We assessed whether application of EGA to carboxyhaemoglobin dilution provides red cell volume (RCV) measurement with accuracy equivalent to that of CO-haemoximetry, with a smaller infusion volume of carbon-monoxide-saturated autologous blood (COB). Method. We assessed the agreement between repeated COHb% measurements by EGA and simultaneous measurement by CO-haemoximetry, using Bland and Altman plot, in healthy subjects and patients with artificially controlled ventilation and no radiological evidence of pulmonary oedema or atelectasis. We assessed the agreement between RCV measurements by EGA with infusion of 20 ml of COB (RCVEGA) and RCV measurements by CO-haemoximetry with infusion of 100 ml of COB (RCVHEM), in healthy subjects., Results: The 'limits of agreement' between COHb% measurement by EGA (1 min average) and CO-haemoximetry were -0.09 and 0.08% in healthy subjects, and -0.11 and 0.09% in patients. Given the resolution of CO-haemoximetry (0.1%), the accuracy of EGA was equivalent to or greater than that of CO-haemoximetry. The 'limits of agreement' between RCVEGA and RCVHEM were -0.14 and 0.15 litre. Given the average resolution of RCVHEM (0.14 litre), the accuracy of RCVEGA was equivalent to that of RCVHEM., Conclusion: EGA provided non-invasive, accurate, continuous, high-resolution COHb% measurements. Applying EGA to carboxyhaemoglobin dilution, we achieved RCV measurements with accuracy equivalent to that of CO-haemoximetry, with one-fifth of the COB infusion volume. However, clinical application of the method is limited to patients with no radiological evidence of pulmonary oedema or atelectasis.

Two conjugates of p-hydroxymethamphetamine (p-OHMA), p-OHMA-glucuronide (p-OHMA-Glu) and p-OHMA-sulfate (p-OHMA-Sul) have been identified in methamphetamine (MA) users' urine by using liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS-MS). The synthesis of p-OHMA-Glu and p-OHMA-Sul, and an LC-MS procedure for the simultaneous determination of MA and its four metabolites, amphetamine (AP), p-OHMA, p-OHMA-Glu and p-OHMA-Sul, in urine have also been established. After deproteinizing urine samples with methanol, LC-MS employing a C(18) semi-micro column with a gradient elution program provided the successful separations and MS determinations of these analytes within 20 min. Based on the established method, p-OHMA-Sul was detected at higher concentrations than p-OHMA-Glu in all of the three urine samples tested. These data suggest that sulfation is a major pathway in the MA phase II metabolism.

Chemokine receptors have attracted a good deal of public attention as important therapeutic targets for many diseases and disorders. In this issue of Chemistry & Biology, Kumar and colleagues propose a new concept of synthetic modular modifications to generate unnatural chemokines, which exhibit high receptor selectivity .

To prove the intake of recently controlled designer drugs, N-benzylpiperazine (BZP) and 1-(3-trifluoromethylphenyl)piperazine (TFMPP), a simple, sensitive and reliable method which allows us to simultaneously detect BZP, TFMPP and their major metabolite in human urine has been established by coupling gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). GC-MS accompanied by trifluoroacetyl (TFA) derivatization and LC-MS analyses were performed after the enzymatic hydrolysis and the solid phase extraction with OASIS HLB, and BZP, TFMPP and their major metabolites, 4'-hydroxy-BZP (p-OH-BZP), 3'-hydroxy-BZP (m-OH-BZP) and 4'-hydroxy-TFMPP (p-OH-TFMPP), have found to be satisfactorily separated on a semi-micro SCX column with acetonitrile-40 mM ammonium acetate buffer (pH 4) (75:25, v/v) as the eluent. The detection limits produced by GC-MS were estimated to be from 50 ng/ml to 1 microg/ml in the scan mode, and from 200 to 500 ng/ml in the selected ion monitoring (SIM) mode. Upon applying the LC-ESI-MS technique, the linear calibration curves were obtained by using the SIM mode for all analytes in the concentration range from 10 ng/ml to 10 microg/ml. The detection limits ranged from 5 to 40 ng/ml in the scan mode, and from 0.2 to 1 ng/ml in the SIM mode. These results indicate the high reliability and sensitivity of the present procedure, and this procedure will be applicable for proof of intake of BZP and TFMPP in forensic toxicology.

Objective: We developed a new apparatus for long-term heart preservation that combines simple immersion with coronary perfusion. In a previous study, we reported that suppression of pro-inflammatory cytokines, such as tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta), improved results after transplantation. In this study, we evaluated whether long-term preservation using our apparatus for continuous coronary perfusion, combined with suppression of pro-inflammatory cytokines, improves donor heart function after transplantation in a canine model., Methods: We used adult mongrel dogs in this study. Coronary vascular beds were washed with University of Wisconsin (UW) solution after arresting hearts with glucose-insulin-potassium solution. The heart was then excised and preserved for 12 hours with a combination of immersion and coronary perfusion using a preservation apparatus. Adult mongrel dogs were divided into 2 groups: the coronary perfusion (CP) group (n = 7) and the FR167653 (FR-CP) group (n = 6). In the CP group, we used a 4 degrees C UW solution for immersion and coronary perfusion. In the FR-CP group, we used a 4 degrees C UW solution supplemented with 20 mg/liter of the anti-inflammatory agent FR167653 for immersion and coronary perfusion. At 2 and at 3 hours after orthotopic transplantation, we compared hemodynamic parameters with pre-operative values in donor animals, with right atrial pressure at 10 mm Hg and with 5 microg/kg/min dopamine infusion. We compared serum concentrations of TNF-alpha from the coronary sinus and compared electron microscopic studies between the 2 groups., Results: Three hours after transplantation, cardiac output (CO), left ventricular pressure (LVP), and -LVdp/dt were significantly greater (p < 0.05) in the FR-CP group than in the CP group (CO, 178% +/- 65% vs 93% +/- 40%; LVP, 115% +/- 22% vs 73% +/-26%; -LVdp/dt, 168% +/- 13% vs 61% +/- 17%, respectively). Electron microscopic studies showed that glycogen was well preserved in the FR-CP group compared with the CP group. Serum concentrations of TNF-alpha were decreased significantly in the FR-CP group compared with the CP group at 3 hours after reperfusion (161 +/- 54 pg/dl vs 642 +/- 636 pg/dl, respectively)., Conclusion: Hemodynamics after transplantation were significantly better in the FR-CP group than in the CP group. The combined preservation method of continuous perfusion and immersion using our apparatus in conjunction with suppression of pro-inflammatory cytokines improves donor heart function after transplantation.

Early-onset sarcoidosis (EOS) and inheritable Blau syndrome (BS) share characteristic clinical features of juvenile-onset systemic granulomatosis syndrome that mainly affects skin, joints, and eyes. However, no direct evidence has been shown for the possible common origin of these 2 diseases. Recent discovery of CARD15 mutations in BS families encouraged us to investigate similar CARD15 mutations in EOS patients. Among 10 EOS cases retrospectively collected in Japan, heterozygous missense mutations were found in 9 cases; 4 showed a 1000C>T (R334W in amino acid change) that has been reported in BS, 4 showed novel 1487A>T (H496L), 1538T>C (M513T), 1813A>C (T605P), and 2010C>A (N670K), and 1 case showed double 1146C>G (D382E)/1834G>A (A612T) mutations on different alleles. All 6 of these variants of CARD15 showed increased basal nuclear factor (NF)-kappaB activity. These findings indicate that the majority of EOS and BS cases share the common genetic etiology of CARD15 mutations that cause constitutive NF-kappaB activation.

Background: This study investigated the effects of a bradykinin B(2) receptor antagonist, FR173657 (FR), on ischemia-reperfusion (I/R) injury in a canine lung transplantation model., Methods: Eighteen pairs of weight-matched dogs were randomly divided into 3 groups. Six pairs were assigned to the FR(D+R) group, in which FR (100 nmol/kg/h) was administered to the transplant donor continuously beginning 30 minutes before ischemia until the onset of ischemia, and FR was administered to the transplant recipient beginning 30 minutes before reperfusion and continuing for 2 hours after reperfusion. Another 6 pairs of dogs were assigned to the FR(R) group, in which FR was administered only to the recipient in the same manner as in the FR(D+R) group. The other pairs were assigned to the control group, in which vehicle alone was administered. Orthotopic left lung transplantation was performed after 12-hour cold storage in Euro-Collins solution. Fifteen minutes after reperfusion, the right pulmonary artery and the right stem bronchus were ligated. The animals were measured for 4 hours after reperfusion for left pulmonary vascular resistance (L-PVR), cardiac output (CO), arterial oxygen pressure (PaO(2)) and alveolar-arterial oxygen pressure difference (A-aD(O(2))). Lung specimens were harvested for measurement of the wet-to-dry lung weight ratio (WDR), histopathologic studies and polymorphonuclear neutrophil (PMN) count., Results: Compared with the control group, PaO(2), A-aDO(2), L-PVR and CO were all significantly (p < 0.05) improved and WDR significantly (p < 0.05) lower in both the FR(D+R) and FR(R) groups. Moreover, in the FR-treated groups, histologic tissue edema was mild, and PMN infiltration was significantly (p < 0.05) reduced., Conclusions: The bradykinin B(2) receptor antagonist, FR173657, ameliorates I/R injury in lung grafts, indicating that protection of lung grafts can be achieved by the administration of FR solely to the transplant recipient.

Objective: The flow cytometric assay was evaluated as a tool for real-time monitoring of human cytomegalovirus (HCMV)-specific cellular immunity in pregnant women., Methods: We screened for HCMV infection in pregnant women in Sapporo, Japan, during the year 2000, by serologic assays, virus isolation from urine, and PCR to detect DNA in cervical swabs. The frequencies of HCMV-specific CD4+ T cells in pregnant women with serum anti-HCMV IgG antibody were detected by intracellular cytokine (ICC), interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) staining., Results: The levels of intracellular cytokines in pregnant women with serum anti-HCMV IgG antibody were significantly higher than those in women without anti-HCMV IgG antibody (P = 0.011 for IFN-gamma and P = 0.023 for TNF-alpha) but lower than those in non-pregnant women with serum anti-HCMV IgG antibody. Frequencies of HCMV-specific CD4+ T cells were higher in infants with symptomatic congenital infection than in infants with asymptomatic perinatal infection., Conclusions: This ICC assay may reflect immunologic activity against HCMV infection in pregnant women with immunosuppressive conditions.

Background: We investigated the effects of briefly perfusing hearts from non-heart-beating donors (NHBDs) with a Celsior solution before cardiac transplantation., Methods: Donor hearts were left in situ for 20 minutes after cardiac arrest was induced by rapid exsanguination. Twelve donor-recipient pairs of mongrel dogs were divided into 2 groups, the simple immersion (SI, n = 6) group and the coronary perfusion (CP, n = 6) group. Both groups underwent coronary flushing with Celsior, after which hearts from the SI group were stored using simple immersion for 4 hours and hearts from the CP group underwent 1 hour of further perfusion followed by storage for 3 hours. Orthotopic transplantation was then performed. We measured cardiac output, end-systolic maximal elastance (E(max)), left ventricular pressure, and rate pressure product 1 and 2 hours after weaning from cardiopulmonary bypass (CPB). Two hours after weaning from CPB, the hearts were harvested for histopathologic study and to determine the percentage of water content., Results: The cardiac output (CO) recovery rate was significantly higher in the CP group than in the SI Group 1 hour after weaning from CPB (p < 0.05). The CO recovery rate, E(max), and rate pressure product were significantly higher and the percentage of water content was significantly lower in the CP group than in the SI Group 2 hours after weaning from CPB (p < 0.05). Histopathologic damage was more severe in the SI group., Conclusions: The results of this study suggest that short-term coronary perfusion with a Celsior solution may be useful for heart transplantation from NHBDs.

Sediment trap experiments were carried out three times from 1999 to 2000, in the western part of the Seto Inland Sea (Suo-Sound), Japan. We investigated both the particulate flux and the composition of chemical substances in the sediment trap samples. Based on the results, we discuss the origin of particulate organic carbon (POC) collected by the sediment traps in a coastal area. Moreover, we purposed to estimate the flux of the portion of the POC that is derived from phytoplankton photosynthesis. The fluxes of POC varied between 677 and 3424 mgC m(-2) d(-1). Significant positive correlations between POC and aluminum (Al) fluxes suggested that these components show almost the same behaviour. The mean value of the Al flux was about eight times higher than that of Al burial rates on the sediment surface. Therefore, it seems that the POC flux observed with the sediment traps was considerably overestimated. Moreover, judging from the fact that Al is a typical terriginous element, it seems that most of the POC collected in the sediment traps derived from the re-suspended surface sediment or sediment transported laterally from shallow flanks such as intertidal mudflats. The fluxes of chlorophyll a (Chl a) were independent of the POC fluxes, and a relatively consistent correlation was found between Chl a abundance in the water column and the Chl a flux. Moreover, surface sediment Chl a content was approximately 100 times lower than that of suspended matter. Therefore, resuspension and terriginous contributions to Chl a collected in sediment traps are likely to be negligible. The POC content in the trap samples varied between 22.4 and 70.7 mg g(-1) dry weight. The variations of POC contents were positively correlated with the Chl a contents: POC(mg g(-1))=76.5 x Chl a(mg g(-1)) + 26.0 (r=0.95, p<0.01, n=9). This result shows that POC contents strongly corresponded with phytoplankton and their debris. It was also considered that the fraction of POC derived from phytoplankton primary production could be estimated as Chl a content times a certain factor. In this study, we estimated the flux of the portion of the POC originating from phytoplankton production by multiplying the Chl a fluxes by 76.5 (the mean POC:Chl a ratio in the trap samples). These values varied between 308 and 758 mgC m(-2) d(-1), and accounted for 35.1+/-21.2% of total POC flux. Although the amount of POC that originates from phytoplankton photosynthesis was a small portion of total POC flux, it seems to be a large portion of potential primary production in the water column.

Background: This study investigated the effects of a bradykinin B(2) receptor antagonist, FR173657 (FR), on pulmonary ischemia-reperfusion (I/R) injury., Methods: Twenty-four mongrel dogs were divided into four groups (n = 6 each). In Groups I, II and III, FR doses of 33, 100 and 300 nmol/kg per hour, respectively, were administered continuously beginning 30 minutes before ischemia and continuing for 2 hours after reperfusion. In Group IV, vehicle alone was administered. Warm ischemia was induced for 3 hours by clamping the left pulmonary artery and veins. Simultaneously, the left stem bronchus was bisected and then anastomosed before reperfusion. Fifteen minutes after reperfusion, the right pulmonary artery and bronchus were ligated. Left pulmonary vascular resistance (L-PVR), cardiac output (CO), arterial oxygen pressure (PaO(2)) and the alveolar - arterial oxygen pressure difference (A-aDO2) were measured for 4 hours after reperfusion. Lung tissue was harvested for wet-to-dry weight ratio (WDR) measurements, histopathologic studies and polymorphonuclear neutrophil (PMN) counts. Serum thromboxane (TX) B(2), 6-keto-prostaglandin (PG) F(1alpha) and leukotriene (LT) B(4) levels were also measured., Results: PaO(2), A-aDO2, L-PVR and CO were significantly (p < 0.05) improved and WDR was significantly (p < 0.05) lower in Groups II and III than in Group IV. Histologic tissue edema was mild, and PMN infiltration was significantly (p < 0.05) reduced in Groups I, II and III compared with Group IV. TXB(2) levels were significantly (p < 0.05) lower in Group II than in Group IV, whereas 6-keto-PGF(1alpha) levels were not significantly different. LTB(4) levels were significantly (p < 0.05) lower in Groups II and III than in Group IV., Conclusions: FR appears to have a protective effect on pulmonary I/R injury stemming from the inhibition of eicosanoid release.

Objective: University of Wisconsin (UW) solution is widely used in organ preservation. Some investigators have reported that high pressure during initial flush with UW solution may induce vasoconstriction and endothelial damage, because of its high potassium content and high viscosity. However, using lower pressure during the initial flush may lead to irregular distribution of the solution and incomplete flushing of blood components from coronary vascular beds. This experimental study evaluated the effects of a range of initial flush pressures during heart procurement, followed by orthotopic transplantation of the graft after 12 hours of preservation., Materials and Methods: Twelve pairs of adult mongrel dogs, weighing 9 to 14 kg, formed the recipient-donor combinations. After determining hemodynamic status by measuring cardiac output, left ventricular pressure (LVP), and maximum positive and negative change in LVP (+/-LVdP/dt), donor hearts were excised. Coronary vascular beds were flushed with 4 degrees C UW solution at a pressure of 60 mm Hg in the low-pressure group (n = 6) and at 120 mm Hg in the high-pressure group (n = 6). After 12 hours of cold preservation, orthotopic transplantation was performed using cardiopulmonary bypass (CPB). The hemodynamics of the transplanted graft were assessed by comparing recovery rates (%) from donor hearts 2 hours after weaning from CPB. Endothelin-1 (ET-1) levels were measured in the blood obtained from the coronary sinus 30 minutes after reperfusion. The transplanted grafts were then harvested for histologic study and measurement of adenosine triphosphate (ATP) content., Results: Cardiac output, LVP, LVdP/dt and myocardial tissue ATP content were significantly better (p < 0.05) in the high-pressure group than in the low-pressure group. We found no significant differences in ET-1 levels between the groups. Transmission electron microscopic findings revealed that degeneration of the mitochondria was less extensive in the high-pressure group than in the low-pressure group. We observed no obvious ultrastructural damage to the endothelial cells in either group., Conclusion: When using UW solution in heart procurement, high pressure is better to completely wash out the blood components and distribute the solution.

We have developed a new surgical technique for the treatment of Tile C-1 type sacroiliac disruption. We tried this procedure first in a cadaveric specimen and then applied it to a clinical case. We used the Texas Scottish Rite Hospital (TSRH) rod and pedicle screw system to insert one screw into the S1 vertebra without using an image intensifier and the other screw into the bone marrow of the ilium from the posterosuperior iliac spine. A straight rod was connected between the two screws by using a manipulator to attempt to reduce and fix the sacroiliac disruption. The combined pubic symphysis diastasis could be simultaneously reduced and fixed by using a plate through another incision, resulting in anatomically correct reconstruction of the pelvic ring. In this procedure, the alignment of the sacroiliac joint can be reversibly and directly changed during reduction and fixation. The sacroiliac joint can be strongly fixed because the screws can be freely inserted into the intact portion of the pelvis and the adjacent lumbar spine, if necessary. Good reduction is obtained because direct compression force is applied to the fracture site. The posterior and anterior procedures can be simultaneously performed under the same lateral position.

Background: In the process of ischemia-reperfusion, inflammatory cytokines and arachidonic acid metabolites are released and followed by tissue damage. FK3311 (FK) is a selective cyclooxygenase-2 inhibitor that inhibits conversion of arachidonic acid into thromboxane A2 or prostaglandin I2. We investigated the effects of FK in canine lung transplantation., Methods: FK3311 was administered in the FK group, and vehicle was injected in the control group. The left lung was orthotopically transplanted after 12-hour preservation in Euro-Collins solution. After reperfusion, the right pulmonary artery and bronchus were ligated, and the animals were observed. Pulmonary gas exchange and hemodynamics were measured, histopathologic damages were investigated, and technetium-99m-labeled albumin scintigraphy was performed. The serum prostanoid levels were also measured., Results: In the FK group, pulmonary gas exchange and hemodynamics were significantly (p < 0.05) better, histologic damage and neutrophil infiltration was reduced, and technetium-99m-albumin accumulation was considerably suppressed. Also, thromboxane B2 was significantly (p < 0.05) lower, but 6-keto-prostaglandin F1alpha was not significantly reduced., Conclusions: FK3311 generates protective effects on lung transplantation by a marked inhibition of thromboxane A2.

Background: Interleukin (IL)-1 and tumor necrosis factor-alpha (TNF-alpha) are recognized as important factors in ischemia-reperfusion (I/R) injury. FR167653 has been characterized as a potent suppressant of IL-1 and TNF-alpha production. We previously reported that FR167653 suppressed the expression of IL-1 beta mRNA after reperfusion and ameliorated pulmonary I/R injury following 3-hour left lung warm ischemia in dogs. The aim of this study was to investigate the effects of FR167653 on I/R injury in a canine left, single, lung transplantation model., Methods: We used 10 pairs of weight-matched dogs. We assigned 5 pairs to the FR group, in which each animal received FR167653 (1 mg/kg/hr) IV from 30 minutes before ischemia until 2 hours after reperfusion; we treated the transplanted lungs with FR167653 after the onset of reperfusion. The others were assigned to the control group. After 8-hour preservation with 4 degrees C Euro-Collins solution, orthotopic left, single, lung transplantation was performed. During a 5-minute clamping test at the right pulmonary artery of each recipient, the left (transplanted) pulmonary arterial pressure (L-PAP), left (transplanted) pulmonary vascular resistance (L-PVR), arterial oxygen pressure (PaO(2)), and alveolar-arterial oxygen pressure difference (A-aDO(2)) were measured. We harvested transplanted lung specimens for histologic study, and we counted polymorphonuclear neutrophils (PMNs), which were identified by staining with naphthol AS-D cholroacetate esterase. Pulmonary perfusion and ventilation scintigraphy (Tc-99m-MAA and Xe-133) were performed. We observed the animals for 3 days after transplantation., Results: The PAP, L-PVR, PaO(2), and A-aDO(2) revealed significantly (p < 0.05) better function in the FR group than in the control group. Histologically, lung edema was milder, and PMN infiltration was significantly (p < 0.05) lower in the FR group than in the control group. Xe-133 and Tc-99m-MAA were widely distributed throughout the graft lung in the FR group. Three-day survival rates in FR and control groups were 60% and 20%, respectively., Conclusions: FR167653 appears to generate a protective effect on I/R injury in lung transplantation in dogs.

Objective: FK409 is the first spontaneous nitric oxide donor to increase plasma guanosine 3':5'-cyclic monophosphate. We designed this study to investigate whether the administration of FK409 during reperfusion ameliorated ischemia-reperfusion injury and enhanced post-transplant graft function in orthotopic heart transplantation following 12-hour cold preservation in a canine model., Methods: We used 10 pairs of adult mongrel dogs, weighing 9.5 to 13.5 kg. Following cardiac arrest using cardioplegia, we washed out the coronary vascular beds with cold University of Wisconsin solution followed by 12-hour preservation. After preservation, we performed orthotopic transplantation. The experimental animals were divided into 2 groups. In the FK group (n = 5), FK409 (5 microg/kg/min) was administered intravenously, beginning 15 minutes before the onset of reperfusion and continuing for 45 minutes after reperfusion. In the control group (n = 5), saline vehicle was administered in the same manner. Two hours after transplantation, we assessed cardiac function, including cardiac output, left ventricular systolic pressure (LVP), and the maximum rates of positive and negative increase of LVP (+/-LV dP/dt) by comparing the recovery rate (%) of the cardiac function of the donor animal. We measured endothelin-1 levels in blood obtained from a catheter inserted into the coronary sinus 30, 60, and 120 minutes after reperfusion., Results: Cardiac output was higher in the FK group than in the control group, but the difference was not significant (p = 0.08). Left ventricular systolic pressure and +/-LV dP/dt were significantly (p < 0.05) higher in the FK group than in the control group. Endothelin-1 levels were significantly (p < 0.05) lower in the FK group than in the control group 30 minutes after reperfusion. Transmission electron microscopy showed that the basal lamina of capillary vessels, glycogen granules, and mitochondrial structure were well-preserved in the FK group., Conclusions: In orthotopic transplantation models, FK409 is effective in ameliorating ischemia-reperfusion injury following preservation and in enhancing post-transplant cardiac function.

Background: Nitric oxide (NO) is known to have beneficial effects in ischemia-reperfusion (I/R) injury through maintaining endothelial integrity, inhibiting leukocyte adhesion and platelet aggregation, and inducing vasodilation. The effect of FK409 (FK), a spontaneous NO donor, was investigated in a canine lung transplantation model., Methods: Ten pairs of weight-matched dogs were used. Five pairs were assigned to the FK group, to which FK (5 microg/kg/min) was administered intravenously from 30 minutes prior to ischemia until the induction of ischemia in the donor, and from 15 minutes prior to reperfusion until 45 minutes after reperfusion in the recipient. The others were assigned to the control group. After 8-hour preservation in 4 degrees C Euro-Collins solution, orthotopic single-lung transplantation was performed. During a 5-minute clamping test of the right pulmonary artery, left pulmonary arterial pressure (L-PAP), left pulmonary vascular resistance (L-PVR), arterial oxygen pressure (PaO(2)), and alveolar-arterial oxygen pressure difference (A-aDO(2)) were measured. The lung specimens were harvested for histologic study, and polymorphonuclear neutrophils (PMNs) were counted. Pulmonary perfusion and ventilation scintigraphy (Tc-99m-MAA and Xe-133) were performed., Results: PAP, L-PVR, PaO(2), and A-aDO(2) revealed significantly (p < 0.05) better function in the FK group than in the control group. Histologically, edema was more mild, and PMN infiltration was significantly (p < 0.05) lower in the FK group than in the control group. Xe-133 and Tc-99m-MAA were widely distributed throughout the graft lung in the FK group. The 2-day survival rate was 100% in the FK group, which was significantly (p < 0.05) better than the rate (40%) in the control group., Conclusions: FK appears to generate a protective effect on I/R injury in lung transplantation.

Progressive burn wound necrosis is an important factor as a cause of delayed healing during clinical therapy of burns. Among the causes of progressive necrosis have been attributed an insufficient blood supply or a dehydration at the zone of stasis just beneath the zone of coagulation. In a previous study evidence was presented that hypernatremia, an osmotic injury, may act to promote progressive tissue or cell death of the superficial dermal wound resulting from a heat injury. To test this hypothesis pathological features of cell death in the second-degree burn wound in the rat with hypernatremia were investigated and evidence for apoptosis in hair follicle cells was observed. Rats in the hypernatremic group were administered 10 ml of hypertonic sodium solution (850 meq 1(-1)) and the control rats were treated with 10 ml of hyponatremic solution (100 meq 1(-1)) to prevent hypernatremia. After 24 h postburn the average incidence of hair follicles (ratio to the normal skin) in the hypernatremic group was 30.1 +/-11.6 per cent and significantly lower when compared with the control group (87.6+/-6.0 per cent). The numbers of hair follicles were studied by haematoxylin and eosin stain, and the apoptotic process was investigated by an immunochemical assay and electron microscopy.

Phagocytic activity is an important function of neutrophils in the host defense against burn wound infection. In a previous report we demonstrated hypernatremic suppression of neutrophils at sodium concentrations comparable to these in the zone of stasis in the burn wound. At this site the osmotic pressure will be elevated as a result of several factors, especially to increased water loss from the burn wound surface. In this study the effect of granulocyte-colony stimulating factor (G-CSF) on phagocytic activity of human neutrophils against hypernatremic suppression is investigated in vitro. In the G-CSF group 150 ng of G-CSF was added to four blood samples (5 ml each). Phagocytic activities of the control group (n = 10) showed 88.3+/-4.1 per cent (means+/-SD) at 140 mmol/l of sodium concentration, 68.2+/-6.9 at 180, 57.6+/-10.1 at 220, and 48.6+/-8.6 at 260 mmol/l. The G-CSF group showed 88.2+/-7.0 at 140, 79.9+/-9.4 at 180, 71.4+/-7.4 at 220, and 56.5+/-14.1 at 260 mmol/l. At 180 and 220 mmol/l significant differences were recognized. Results of this study suggest a favourable effect of G-CSF on suppressed neutrophils under the hypernatremic conditions.

The influence of hypernatremia on progressive burn wound necrosis was evaluated in a second-degree burn model. The depth of the burned tissue sloughed off was measured in a comparative study in the rat (contact burn injury with a metal plate, 70 degrees C for 5 s). Rats in the hypernatremic group were treated with an intraperitoneal fluid injection of hypernatremic saline (10 ml, 850 mEq/l). Control rats were injected with hyponatremic saline (10 ml, 100 mEq/l). On the fourth postburn day specimens were harvested and compared. The greatest depth of leukocytic infiltration (percent of total dermal thickness) was measured. The average depths were significantly different. In zone 1 (from the normal skin edge to 5 mm inside the wound) the average depth in the hypernatremic group was 38.0 +/- 9.3 percent of the dermal depth and in the control group 9.5 +/- 1.2 percent. In zone II (from 5 to 10 mm inside the wound) the hypernatremic group sloughed off at 64.0 +/- 11.8 percent and the control group at 12.5 +/- 2.1 percent. The hypernatremic group showed a deeper demarcating borderline of leukocytic infiltration than the control rats. The wound depth progression may be caused by an osmotic injury.

Phagocytic and bactericidal activities are important functions of the human polymorphonuclear leucocytes (PMNL) as the host defence against burn wound infection. The zone of stasis, just below the zone of coagulation, is a site of interaction between invading bacteria and PMNL. For many reasons the osmotic pressure at this site is elevated. In this study we evaluated the in vitro phagocytic and bactericidal activities of PMNL under hypernatremic conditions. At a sodium concentration of 180 mEq/l, phagocytic activity was suppressed. Although on average superoxide production was maintained within the normal range, it was suppressed in three of the ten cases studied. We conclude that the hypernatremic condition may weaken local defence against burn wound infection at the zone of stasis and may be a risk factor for burn wound sepsis.