Ontogenetic transition from aquatic to amphibious life in the mudskipper, Periophthalmus modestus.

Vertebrates first emerged from water to land in the Paleozoic. Our understanding about the process has been steadily refined through paleontological studies, although the soft-body traits and behavior of these early animals remain poorly known. Mudskippers, extant amphibious gobies, could give insight into this question. This study reports on the ontogenetic transition from water to land of the mudskipper Periophthalmus modestus under laboratory conditions. After about 30 days after hatching (dah), the fish gradually changed their preference from water to an artificial shore and then to land. After about five days of periodic volitional emersion, the fish became able to propel themselves on land using the pectoral fins and after a further 13 days they began feeding on land. During the transition, the head morphology altered to suit for terrestrial existence. Tissue contents of triiodothyronine (T3) and thyroxine (T4) sharply increased at 30 dah. Forced underwater confinement of larvae at the last pelagic stage (27-29 dah) for 40-42 days resulted in no statistically significant difference in survival or gross morphology of the body and the gills. Growth was slightly stimulated. Our results show that mudskippers emerge on land with little morphological alteration during ontogenesis, much less than the changes observed for amphibians, and that emersion was not indispensable for survival or growth under our laboratory conditions. Further analysis of how and why mudskippers make their way across the water's edge will shed valuable light on what morphological, behavioral and physiological traits were needed for, and what environmental conditions may have driven the earliest steps of the water-to-land transition in ancient fishes.

Mutations of the KLF1 gene detected in Japanese with the In(Lu) phenotype.

BACKGROUND: In(Lu) is characterized by a reduced expression of antigens in the Lutheran blood group system as well as other blood group antigens. Mutations of the erythroid transcription factor, KLF1, have been reported to cause the In(Lu) phenotype, and we investigated Japanese In(Lu) to estimate the prevalence of the phenotype and KLF1 polymorphism. STUDY DESIGN AND METHODS: Blood samples were screened by monoclonal anti-CD44 and the In(Lu) phenotype was confirmed by tube tests including adsorption and elution tests using anti-Lua and anti-Lub . KLF1, LU, and A4GALT genes were analyzed by polymerase chain reaction and sequencing. RESULTS: We identified 100 of 481,322 blood donors (0.02%), and the previously characterized 20 donors, who had the In(Lu) phenotype with the LUB/LUB genotype. A total of 100 of the 120 In(Lu) individuals had mutant KLF1 alleles, and we identified 13 known and 21 novel alleles. The mutant KLF1 alleles with c.947G>A (p.Cys316Tyr), c.862A>G (p.Lys288Glu), or c.968C>G (p.Ser323Trp) were major in the In(Lu) individuals. The P1 antigen of 29 In(Lu) (two P1 /P1 , 27 P1 /P2 ) showed significantly weakened expression by hemagglutination. CONCLUSIONS: The prevalence of the In(Lu) phenotype in the Japanese population was 0.02%, and we identified 13 known and 21 novel KLF1 alleles. The KLF1 mutations cause the reduced expression of the P1 antigen.

Fatal warm autoimmune hemolytic anemia in a child due to IgM-type autoantibodies.

Herein is described a case of immunoglobulin M (IgM) warm autoimmune hemolytic anemia (AIHA) in a child who consequently died within 3 days of clinical onset. A previously healthy 11-year-old boy presented with fever, anemia, jaundice, and deteriorating consciousness. On direct agglutination test against group O red blood cells, agglutination was seen even at 37°C in saline, which was abolished on dithiothreitol treatment of the serum, indicating that the responsible autoantibody was IgM and had a warm-reactive capacity. A diagnosis of IgM warm AIHA was therefore made. Hemagglutination in the visceral capillaries was considered as the direct cause of organ dysfunction. The patient died due to respiratory failure. IgM warm AIHA is a very severe condition that is difficult to reverse in an advanced state. Both prompt, definite diagnosis and intervention are therefore vital to prevent severe multi-organ dysfunction in cases of IgM warm AIHA.

[Current status of the treatment strategy for life-threatening hemorrhage in hospitals accredited by the Japanese Society of Anesthesiologists (JSA), and practical use of "Guidelines for Action Against Intraoperative Critical Hemorrhage"].

The JSA and the Japanese Society of Blood Transfusion and Cell Therapy established "Guidelines for Action Against Intraoperative Critical Hemorrhage" in 2007. This guideline shows practical strategies for managing life-threatening hemorrhage. From 2007 to 2009, we conducted the annual survey with transfusion services of 384 hospitals accredited by the JSA. From the results of these surveys, we proposed some new strategies for managing life-threatening hemorrhage as follows: 1) It is necessary to establish a documentation of institutional procedures for urgent transfusion practices including an emergency code for blood requirement, according to "Guidelines for Action Against Intraoperative Critical Hemorrhage". 2) The simulation exercise according to the documentation of institutional emergency transfusion procedures should be held. 3) On the occurrence of a life-threatening hemorrhage, we should make an appropriate decision to use uncrossmatched ABO-matched blood components and/or ABO-mismatched compatible blood components, to save the patient's life. 4) Though it is well known that the post-transfusion graft versus host disease (PT-GVHD) has poor prognosis and irradiation of blood products is essential for avoiding the PT-GVHD, in some hospitals, non-irradiated blood components were used in situation of critical hemorrhage. We recommend that irradiated blood components should be used if possible in resuscitating a patient with critical hemorrhage.

[Advancement in the preparation of red cell concentrates and platelet concentrates].

Both Japan Society of Blood Transfusion and Cell Therapy and Japan Society of Anesthesiologists have made a "Guideline of Management at Critical Bleeding in the Operating Room" in 2007. Since 2008, Japan Red Cross Blood Center (JRC) introduced leuko-reduction filter and diversion technique to prevent bacterial contamination. This improvement can easily introduce ABO compatible transfusion at critical situation. We proved the safety of transitional anti-A, anti-B antibody at compatible transfusion. 1) anti-A, anti-B antibody of leuko-reduced red cell concentrates (RCC-LR): Due to 90% plasma removal by centrifugation, antibody titer of anti-A, anti-B antibodies in RCC-LR is less than 2 times dilution. This level of antibodies does not cause hemolysis when more than 100 compatible ABO mismatch RCC-LR bags transfused at once. 2) Neutralization of ant-A, anti-B antibody due to A, B substance containing patient's plasma: ABO blood type is composed by glucan antigen, and it is contained in the plasma, saliva and many organs. When ABO incompatible transfusion has been done, transitional anti-A, anti-B antibodies are neutralized by A, B substances. Our experiment showed neutralizing titer 64 times equal volume of ABO incompatible plasma. 3) Anti-A, anti-B antibody titer of Fresh Frozen Plasma (FFP) and Platelet Concentrates (PC): With ABO mismatch transfusion, FFP and PC do not contain incompatible red cells at all. Therefore, the risk of hemolysis will be caused by transitional anti-A, anti-B antibodies. Both with PC and FFP, anti-A, anti-B antibodies keep normal level (average: 16 times dilution). The safe volume of ABO mismatch PC and FFP administration has limited within 3 liters. When such mismatch transfusion necessarily performed, hydration therapy to protect kidney function should be applied immediately after hemostasis. 4) Red Cell Volume in a PC bag: PC in Japan have processed by single donor apheresis alone since 2004. Our results showed that each PC bag contains less than 5 mm(-3) of RBCs. If this level of RBCs caused hemolysis in ABO mismatch patient, it is too small to cause DIC or renal failure.

[Emergency compatible, ABO-different blood group transfusion in the operating theater: a survey in regional Japanese Society of Anesthesiologists-certified training hospitals in 2006].

BACKGROUND: In Japan, emergency blood transfusion practices including ABO-compatible, different blood group transfusion and uncross-matched, ABO-identical blood group transfusion are very limited possibly due to adherence to identical blood transfusion as well as fear of hemolytic reactions due to anti-A, anti-B, anti-RhD and unexpected antibodies. Purpose of the study is to examine the incidence of hemolytic reactions due to compatible, ABO-different blood group transfusion. METHODS: We conducted a questionnaire survey regarding emergency compatible, different blood group transfusion in the operating theater in hospitals with more than 500 beds among those with an accredited Department of Anesthesiology regarded as regional hospitals. Of 384 institutions, 247 responded to the questionnaire. During the year 2006, compatible, ABO-different blood group transfusion was reported in 112 patients from 32 hospitals, among which 105 patients in 26 hospitals were available for further analysis. RESULTS: Compatible red cell concentrate (RCC), fresh frozen plasma (FFP), and platelet concentrate (PC) were transfused in 23, 10, and 83 patients, respectively. Total amount of compatible RCC, FFP, and PC were 232, 162, and 1,679 units, respectively. In patients who were transfused with compatible RCC, two patients had unexpected antibodies. Overall mortality rate within the 30th post-operative day was 23%. In 80 patients, in whom only PCs were used as compatible blood products, blood loss was 86 +/- 85 ml x kg(-1), 65% of patients underwent cardiovascular surgery, and mortality was 11%, implying that compatible PC was transfused mainly to avoid hemorrhagic diathesis in cardiovascular patients. In 64 patients with blood type of A, B, or AB, who underwent compatible PC transfusion, type O PC, incompatible blood products, were transfused in 9 patients. In 21 patients, in whom only RCCs were used as compatible blood products, blood loss was 206 +/- 224 ml x kg(-1), and mortality was 57%. Therefore, compatible RCCs were transfused mainly to avoid life-threatening events. Uncross-matched, ABO-identical RCC transfusion was performed only in 29% of patients among these 21 patients. Transfusion-related hemolytic reactions were not reported in all 104 patients available for this analysis. CONCLUSIONS: Although the patient number was small, the finding that there were no hemolytic reactions might promote emergency blood transfusion practices in Japan. High mortality rate and a low rate of uncross-matched, ABO-identical RCC transfusion in patients with compatible RCC transfusion suggest that promoting emergency blood transfusion practices might reduce mortality rate due to massive hemorrhage in the operating theater.

[Present status of preparatory measures for massive hemorrhage and emergency blood transfusion in regional hospitals with an accredited department of anesthesiology in 2006].

BACKGROUND: Annual surveys conducted by the Japanese Society of Anesthesiologists repeatedly show that hemorrhage is the leading cause of life-threatening events in the operating room. METHODS: We performed a questionnaire survey regarding the present status of critical hemorrhage/ blood transfusion occurring in the operating room on an institutional scale and individual blood transfusion management in cases of massive hemorrhage (> or = 5,000 ml) in hospitals with > or = 500 beds and those with an accredited Department of Anesthesiology regarded as regional hospitals. RESULTS: Of 384 institutions, 247 responded to the questionnaire (response rate: 64%), and 692,241 cases managed by anesthesiologists in 2006 were registered. There were 2,657 cases of massive hemorrhage above the circulating blood volume in the operating room, and 404 of them were critical. Thus, the number of cases of massive hemorrhage was 6.6 times that of cases of critical events due to hemorrhage. In the survey of individual cases of massive hemorrhage (> or = 5,000 ml), 1,257 cases were registered in 2006, of whom 196 cases (15.6%) died within 30 post-operative days and 160 cases (12.7%) had some sequelae. The amount of transfused red blood cell concentrate was 25.2 +/- 24.2 units. The amount of red blood cell concentrates stocked for emergency was 12.7 +/- 10.1 units for blood group A, 9.7 +/- 7.3 units for group B, 11.9 +/- 9.6 group AB, and 11.3 +/- 11.0 for group O. Therefore, for those other than group O cases, 21-46 units of red blood cell concentrates seemed to be available in the hospital. The survey of individual cases showed uncross-matched, same blood group transfusion and compatible, different blood group transfusion were performed in only 8.2% and 4.3%, respectively. The lowest hemoglobin concentration was below 5 g x dl(-1) in 16.7% of the cases, but uncross-matched, same blood group transfusion was performed only in 19.0% and compatible, different blood group red cell concentrate transfusion in 5.2%. Even in cases who required cardiac massage, uncross-matched, same blood group transfusion was performed only in 17.1% and compatible, different blood group red cell concentrate transfusion in 8.5%. Intraoperative blood salvage was performed in only 5.7% in cases who underwent non-cardiac surgery. The "Guidelines for the Management of Critical Hemorrhage" proposed in 2007 or the manuals for in-hospital emergency blood transfusion were insufficiently recognized, even by anesthesiologists, and rarely known by surgeons. There were no such manuals in more than 60% of the institutions. CONCLUSIONS: Undertransfusion may occur in 16.7-28.3% of cases of massive hemorrhage in the operating room, and the rate of emergency blood transfusion was much lower than this percentage. To avoid operation-associated deaths from hemorrhage, the improvement of hospital systems for emergency blood transfusion, including the active use of intraoperative blood salvage, should be promoted.

Large volume autologous plasma predonation and retransfusion in orthopedic surgical patients.

BACKGROUND: Intraoperative washed autologous transfusion of the scavenged blood can reduce the deterioration of anemia, even during the operation with a comparatively large blood loss. On the other hand, plasma level can not be collected by this system. The preoperative donation and perioperative retransfusion of autologous plasma may reduce the plasma dilution. PURPOSE: The influence of a large volume plasma predonation and perioperative retransfusion on the plasma protein level was investigated. METHODS: Thirteen patients (63.2 +/- 13.2 yr, 70.3 +/- 12.1 kg) were examined regarding their serum protein (SP), IgG, coagulation systems, colloid osmotic pressure (COP), blood cell count before, just after, 2 h after and 7 days after the donation of 900 ml plasma by plasmapheresis with a simultaneous volume replacement. Twenty surgical patients (52.8 +/- 17.3 yr, 72.6 +/- 16.6 kg, the mean predonated autologous plasma: 2100 ml) with intra- and postoperative retransfusion of autologous plasma were examined perioperatively for SP, IgG, coagulation systems and COP. These parameters were compared with that of the predonated plasma. RESULTS: All data including SP, coagulation and COP, with the exception of IgG, completely recovered within 7 days after preoperative plasmapheresis. Perioperatively, autologous washed blood transfusion system was used. The retransfused volume of autologous predonated plasma was 1740 ml on average. Although about 41 of blood on average was lost perioperatively, only one patient out of 20 patients had to be administered homologous red blood cell transfusion. The levels of most parameters, except for COP, constantly recovered in accordance with the autologous plasma transfusion. Differences in the patterns of improvement were also observed between the parameters. CONCLUSION: A 900 ml plasma predonation can therefore be safely performed with an interval of not less than a week between the last donation and the operation. Autologous plasma retransfusion is thus considered to improve the protein levels.

Granulocytapheresis in patients with refractory ocular Behcet's disease.

Intraocular inflammation (uveoretinitis) is one major complication of Behcet's disease (BD) and responds poorly to drug therapy. This open prospective study was to assess the efficacy of selective granulocytapheresis in patients with refractory uveoretinitis of BD. Fourteen patients aged 20-56 years were treated. Granulocytapheresis was done with an Adacolumn filled with cellulose acetate leucocyte carries or beads that adsorb granulocytes and monocytes from the blood in the column. Each patient received 5 Adacolumn sessions at one session/week over 5 consecutive weeks. The study was designed to allow each patient to serve as his or her own control. The total numbers of ocular attacks (OA) were monitored for 6 months before and after 5 Adacolumn sessions. The number of OA (mean +/- SD) per patient for the 6 months before Adacolumn was 4.21 +/- 1.6 and for the 6 months post Adacolumn was 2.93 +/- 1.39 ( P = 0.0275). Nine patients (64%) improved and 5 did not change or worsened. Further, for a sub-group (n = 7) with duration of BD > or =5 years, the number of OA were 4.71 +/- 1.89 for the first 6 months and 2.29 +/- 1.38 for the second 6 months ( P = 0.0054). The corresponding values for a sub-group (n = 7) with duration of BD<5 years were 3.71 +/- 1.25 and 3.57 +/- 1.13, indicating that patients with long duration of BD are better responders. We conclude that granulocytapheresis might be effective and safe for patients with refractory ocular BD. Further studies are necessary to fully evaluate the clinical efficacy of granulocytapheresis for BD.

Mobilization of human lymphoid progenitors after treatment with granulocyte colony-stimulating factor.

Hemopoietic stem and progenitor cells ordinarily residing within bone marrow are released into the circulation following G-CSF administration. Such mobilization has a great clinical impact on hemopoietic stem cell transplantation. Underlying mechanisms are incompletely understood, but may involve G-CSF-induced modulation of chemokines, adhesion molecules, and proteolytic enzymes. We studied G-CSF-induced mobilization of CD34+ CD10+ CD19- Lin- and CD34+ CD10+ CD19+ Lin- cells (early B and pro-B cells, respectively). These mobilized lymphoid populations could differentiate only into B/NK cells or B cells equivalent to their marrow counterparts. Mobilized lymphoid progenitors expressed lymphoid- but not myeloid-related genes including the G-CSF receptor gene, and displayed the same pattern of Ig rearrangement status as their bone marrow counterparts. Decreased expression of VLA-4 and CXCR-4 on mobilized lymphoid progenitors as well as multipotent and myeloid progenitors indicated lineage-independent involvement of these molecules in G-CSF-induced mobilization. The results suggest that by acting through multiple trans-acting signals, G-CSF can mobilize not only myeloid-committed populations but a variety of resident marrow cell populations including lymphoid progenitors.

[Allogenic blood transfusion for surgery].

Blood transfusion against surgical patients is mainly as a replacement therapy for intra-operative bleeding. Transfusion trigger depends on to maintain 12 ml/kg/min oxygen carrying capacity. Oxygen consumption depends on oxygen carrying capacity which multiplicity hemoglobin concentration and cardiac output. The government of United States decided the transfusion trigger of surgical patients at hemoglobin 7.0 g/dl except patients who have cardiac problem at 1988. The indication of plasma and platelet transfusion is limited in surgical field. Because bleeding tendency is usually a contra-indication of surgery itself. Dilution coagulopathy due to massive bleeding exceed one blood volume is the main indication of plasma and platelet transfusion. It is necessary to administrate fresh frozen plasma for replenishing coagulation function with 30% of one body plasma volume when prothrombin time prolong more than 16 seconds. Platelet transfusion is effective for hemostasis in case of massive bleeding which exceed 1.5 blood volume and also peripheral blood platelet count indicate lower than 50,000/mm3.

Conditioning with targeted busulfan for autologous peripheral blood stem cells transplantation for acute myelogenous leukemia in an XYY male.

We report herein a 19-year-old Japanese male with XYY syndrome who developed acute myelogenous leukemia. During three courses of cytotoxic chemotherapy, he suffered repeated hepatic and renal insufficiencies, possibly related to latent dysfunction from the XYY syndrome. The patient was treated with granulocyte colony-stimulating factor combined with etoposide, cytarabine, and busulfan (the latter adjusted to a targeting dose) followed by autologous peripheral blood stem cell transplantation. He had no severe regimen-related toxicities and is now free of leukemia.

Nonmyeloablative allogeneic bone marrow transplantation for treatment of myelodysplastic syndrome complicated by recent intracerebral hemorrhage.

A patient with intracerebral hemorrhage is considered ineligible for hematopoietic stem cell transplantation (HSCT). We report a 49-year-old woman with myelodysplastic syndrome (MDS) complicated by refractoriness to platelet transfusion and intracerebral hemorrhage, who underwent allogeneic bone marrow transplantation from an HLA-identical unrelated male donor. Nine days before the scheduled transplantation, she developed dysarthria and right hemiparesis; computed tomography (CT) of the brain disclosed an acute hematoma in the left parietal lobe exceeding 3 cm in diameter. She underwent conditioning with reduced-intensity, including fludarabine (30 mg/m2/day on days -8 to -3), busulfan (4 mg/kg/day on days -6 and -5), and total body irradiation (4 Gy on day -2). Two weeks after transplantation, dysarthria and right hemiparesis began to resolve, and CT showed spontaneous resolution of the hematoma. Simultaneously, engraftment was confirmed. Thus, recent stroke may be not an absolute contraindication for HSCT.

Effect of immunotherapy in myelitis with atopic diathesis.

OBJECTIVE: A recent nationwide survey of myelitis with atopic diathesis in Japan disclosed that the disease frequently shows a chronic persistent course. A neuropathological study of the spinal cord also revealed chronic active inflammation. Since the effects of various immunotherapies have not been studied extensively in this condition, we evaluated the efficacies of various immunotherapies in patients with myelitis with atopic diathesis. PATIENTS AND METHODS: Forty-two treatments in 26 patients with myelitis with atopic diathesis were retrospectively analyzed. One of the following therapies was administered: (1) corticosteroids (CS) (pulse therapy followed by oral administration with gradual tapering); (2) intravenous immunoglobulin (IVIG) (400 mg/kg/day for 5 consecutive days); (3) plasma exchanges (PE); or (4) PE followed by IVIG or CS (PE+IVIG/CS). The therapeutic efficacies were evaluated by thorough neurological examination and laboratory tests including MRI, somatosensory evoked potentials (SEPs) and motor evoked potentials (MEPs). RESULTS: Objective neurological findings improved in 89% of the PE group and 90% of the PE+IVIG/CS group, compared with only 72% of the CS and 60% of the IVIG groups. Improvement determined by laboratory tests was seen in 57% of the PE and 57% of the PE+IVIG/CS groups, compared with only 15% of the CS and none of the IVIG groups. Thus, the improvement rate determined by laboratory tests was significantly greater for therapies including PE than for those without PE (p=0.0187). CONCLUSIONS: These data suggest that immunotherapy is effective in myelitis with atopic diathesis despite a chronic persistent course, and that PE is the most beneficial immunotherapy.

[Nonmyeloablative allogeneic stem cell transplantation for a 70-year-old woman with relapsed acute myelogenous leukemia].

We describe a 70-year-old woman with acute myelogenous leukemia with t(8;21) in the first relapse who underwent nonmyeloablative transplantation with conditioning of fludarabine and low-dose total body irradiation (2Gy). Myelosuppression was very mild, and the patient developed transient grade I renal and hepatic toxicities. Complete chimerism was achieved on day 120. The level of the AML1/MTG8 fusion gene in bone marrow decreased to an undetectable level on day 56 and the patient is alive and in complete remission with a follow-up at day 450. This transplant regimen might be well tolerated even by the elderly patients and bring a durable remission.

Successful treatment with nonmyeloablative allogeneic hematopoietic stem cell transplantation in a patient with acute myeloid leukemia complicated with pulmonary infection.

We describe the case of a 48-year-old man with acute myeloid leukemia complicated with pulmonary infection that was successfully treated by nonmyeloablative allogeneic peripheral blood stem cell transplantation with conditioning by low-dose total body irradiation and fludarabine. The disease was diagnosed immunophenotypically as myeloid/natural killer cell precursor acute leukemia. After two courses of induction therapy, complete remission was achieved. However, the patient developed pneumonia from prolonged severe neutropenia. Nonmyeloablative allogeneic transplantation was performed because of the active pulmonary infection and the patient's poor performance status. Myelosuppression after transplantation was mild, and the pulmonary infiltration was well controlled during the course of treatment. At the time of this report the patient was an outpatient in our clinic, and on day 500, his disease was in remission with well-controlled chronic graft-versus-host disease. Nonmyeloablative transplantation may provide a new therapeutic strategy for treating patients with active infection who cannot tolerate conventional transplantation with high-dose chemoradiotherapy.