Less maintenance immunosuppression in lung transplantation following hematopoietic stem cell transplantation from the same living donor.

Living-donor lobar lung transplantation (LDLLT) is one of the final options for saving patients with pulmonary complications after hematopoietic stem cell transplantation (HSCT). We retrospectively investigated 19 patients who had undergone LDLLT after HSCT in Japan. Eight patients underwent LDLLT after HSCT in which one of the donors was the same living donor as in HSCT (SD group), while 11 received LDLLT from relatives who were not the HSCT donors (non-SD group). In the SD group, three patients underwent single LDLLT. The 5-year survival rate was 100% and 58% in the SD and non-SD groups, respectively. In the SD group, postoperative immunosuppression was significantly lower than in the non-SD group. Two patients died of infection and one died of post-transplant lymphoproliferative disease (PTLD) in the non-SD group, while only one patient died of PTLD 7 years after LDLLT in the SD group. Hematologic malignancy relapsed in two patients in the non-SD group. For the three single LDLLTs in the SD group, immunosuppression was carefully tapered. In our study, LDLLT involving the same donor as for HSCT appeared to have advantages related to lower immunosuppression compared to LDLLT from relatives who were not the HSCT donors.

Dynamic instability of central airways and peripheral airspace in rat lungs perfused with cold preservation solutions.

BACKGROUND/AIMS: For lung preservation, one of two types of solutions is commonly employed: Euro-Collins (EC) or low potassium dextran glucose (LPDG). These two solutions have been compared regarding biological, morphometrical and physiological outcomes in many experiments. However, the dynamic mechanics of perfused lung are not well understood because the dynamic characteristics cannot be assessed under static conditions; hence, the primary goal of the present study was to assess this in perfused rat lungs during the preservation period, comparing EC with LPDG at 0 or 9 h at 4°C. METHODS: Lung impedance was measured using a forced oscillation technique. Lung resistance and elastance values were obtained by the fast Fourier transform algorithm. The instability of central airways and heterogeneity of ventilation were estimated. RESULTS: In the EC group, airway resistance and instability were high after perfusion, and the lung elastance was high and more heterogeneous after cold storage. In contrast, those parameters were stable in the LPDG group during cold storage. CONCLUSION: Such dynamic stability might facilitate the handling of lung grafts and eliminate injurious cyclic ventilation stress after reperfusion. Thus, we conclude that the impedance frequency characteristic represents a novel informative parameter for investigating lung preservation techniques.

Potential for pulmonary protection by nebulized milrinone during warm ischemia.

OBJECTIVE: A method to compensate for the donor shortage may be the utilization of donation after cardiac death. The control of lung injury against warm ischemia is crucial in manipulating donors after cardiac death. Nebulization is a simple and feasible drug delivery route after cardiac death. Herein we have examined the potential effect of nebulized milrinone, a phosphodiesterase III inhibitor, on pulmonary warm ischemia. MATERIALS AND METHODS: Deeply anesthetized rats were euthanized by exsanguination. Lungs were exposed to warm ischemia with ventilation up to 2 hours. Milrinone was nebulized for 10 minutes at the beginning of warm ischemia (n = 5). In the control group (n = 5), normal saline was nebulized for the same time. At given intervals, the lungs were partially resected to measure adenine nucleotide and cyclic adenosine monophosphate levels. RESULTS: In both groups, lung tissue cyclic adenosine monophosphate levels decreased significantly at 2 hours after warm ischemia; however, there was no significant difference between the groups. Lung tissue adenosine triphosphate levels significantly decreased at 2 hours after ischemia in the control group, while they did not drop up to 2 hours in the milrinone group. Further, lung tissue adenosine triphosphate levels at 2 hours after ischemia were higher in the milrinone group than the control group. CONCLUSIONS: Our results confirmed that milrinone nebulization during warm ischemia maintained lung tissue adenosine triphosphate levels. Therefore, milrinone nebulization may have potential for lung protection against warm ischemia.

[Mediastinal aberrant goiter; report of a case].

A 54-year-old woman was admitted to our hospital because of an abnormal shadow on chest X-ray. Chest computed tomography (CT) scan and magnetic resonance imaging (MRI) demonstrated an anterior mediastinal tumor. The tumor was resected completely through a median sternotomy. The tumor was dissected successfully from the surrounding vessels in spite of the heavy adhesion to them. The blood supply of the tumor was from a branch of the brachiocephalic artery. The tumor was 9 x 8 x 3 cm in size, and was diagnosed as an aberrant mediastinal goiter since it showed no communication to the thyroid gland. An aberrant mediastinal goiter is a quite rare entity of diseases and its removal through the neck would result in uncontrolled blood loss because its blood supply usually derives from intrathoracic vessels.

[Viral infection after lung transplantation].

We experienced 3 cases of viral infections after lung transplantation. Case 1: Fifty-two-year-old male with pulmonary emphysema underwent left single lung transplantation from a cadaveric donor. Three months after transplantation he presented Epstein-Barr virus (EBV) viremia, resulting in multiple lymphadenopathy. Biopsy showed post-transplant lymphproliferative disorder, and he was treated successfully with rituximab. He is well without recurrence around 1 and a half years after treatment. Case 2: Thitry-eight-year-old male with pulmonary emphysema underwent double lung transplantation from a cadaveric donor. Four months after transplantation he showed multiple nodules in both lungs. Percutaneous biopsy showed post-transplant lymphproliferative disorder, and he was treated successfully with rituximab. He is well without recurrence more than 2 years after treatment. Case 3 : Twenty-four-year-old woman with lymphangioleiomyomatosis underwent living-related bilateral lobar lung transplantation. Three months after lung transplantation she presented cytomegalovirus viremia. Since it proved to be ganciclovir-resistant cytomegalovirus infection, she was treated with foscarnet successfully. She is well without recurrence about 2 and a half years after treatment.

Recurrent lymphangioleiomyomatosis after living-donor lobar lung transplantation.

Living-donor lobar lung transplantation (LDLLT) has been applied to patients with various end-stage lung diseases. The recurrence of pulmonary lymphangioleiomyomatosis (LAM) after lung transplantation has been rarely reported. Herein, we report a case of recurrent pulmonary LAM after LDLLT. A 24-year-old woman presented with pneumothorax and infiltrates in the left lung 1 year after bilateral LDLLT for LAM. These symptoms and radiologic findings occurred repeatedly and then improved quickly. Thereafter, computed tomography of the chest revealed a tiny emphysematous change of the subpleural region in the left lung, which was exacerbated gradually and was finally diagnosed as LAM recurrence by transbronchial lung biopsy. In previous reports of LAM recurrence, the diagnosis was made at the time of autopsy. This is also the first reported case diagnosed early, that is, when the patient was alive and her allograft had not deteriorated badly.

Urethral transitional cell carcinoma in a cat.

A 15-year-old, male neutered cat was referred for investigation of dysuria. A retrograde urethrography was performed which showed two space-occupying masses within the lumen of the mid-to-proximal urethra. Exploratory coeliotomy revealed two urethral masses. Segmental urethrectomy was performed to resect the mass, and the lower urinary tract was reconstructed by vesico-urethral anastomosis. Histopathology showed the mass to be a transitional cell carcinoma with incomplete surgical margins. Tumour regrowth was suspected when dysuria was found approximately 318 days after surgery. Clinical signs were palliated by radiation using weekly fractions of 6 Gy for three weeks. The cat died of unknown causes 386 days postoperatively.

Topical formulations and wound healing applications of chitosan.

Chitosan is being used as a wound-healing accelerator in veterinary medicine. To our knowledge, chitosan enhances the functions of inflammatory cells such as polymorphonuclear leukocytes (PMN) (phagocytosis, production of osteopontin and leukotriene B4), macrophages (phagocytosis, production of interleukin (IL)-1, transforming growth factor beta 1 and platelet derived growth factor), and fibroblasts (production of IL-8). As a result, chitosan promotes granulation and organization, therefore chitosan is beneficial for the large open wounds of animals. However, there are some reported complications of chitosan application. Firstly, chitosan causes lethal pneumonia in dogs which are given a high dose of chitosan. In spite of application of chitosan to various species, this finding is observed only in dogs. Secondly, intratumor injection of chitosan on mice bearing tumor increases the rate of metastasis and tumor growth. Therefore, it is important to consider these effects of chitosan, prior to drug delivery.

Physiological production of singlet molecular oxygen in the myeloperoxidase-H2O2-chloride system.

The putative role of singlet oxygen (1O2) in the respiratory burst of neutrophils has remained elusive due to the lack of reliable means to study its quantitative production. To measure 1O2 directly from biological or chemical reactions in the near infrared region, we have developed a highly sensitive detection system which employs two InGaAs/InP pin photodiodes incorporated with a dual charge integrating amplifier circuit. Using this detection system, we detected light emission derived from a myeloperoxidase (MPO)-mediated reaction in physiological conditions: pH 7.4, 1-30 nM MPO, 10-100 microM H2O2 and 100-130 mM CI in place of Br without the use of deuterium oxide. The MNPO-H2O2-CI(-) system exhibited a single emission peak at 1.27 microm with a spectral distribution identical to that of delta singlet oxygen. Our results suggest physiological production of 1O2 in the MPO-H2O2-CI(-) system at an intravacuolar neutral pH. The MPO-mediated generation of 1O2, which may have an important role in host defense mechanisms, is discussed in connection with previous results.

A prefecture-wide survey of mumps meningitis associated with measles, mumps and rubella vaccine.

A survey of untoward reactions, especially central nervous system reactions, after the administration of a newly introduced measles, mumps and rubella (MMR) vaccine in Gunma Prefecture, Japan, was initiated soon after 4 patients were hospitalized for aseptic meningitis. Thirty-five, 6 and 2 children developed meningitis, convulsive disorders and parotitis, respectively, within 2 months after MMR vaccination during the 8-month period extending from April to November, 1989. The time lag between MMR vaccination and meningitis ranged from 14 to 28 days in the 35 cases of meningitis. Mumps virus, isolated from the cerebrospinal fluid in 13 patients with aseptic meningitis, was characterized by determination of the nucleotide sequences of the P gene as mumps vaccine strain. The incidence of aseptic meningitis with positive mumps vaccine virus was estimated to be 0.11% (0.3% as a whole) during the 8 months from April to November and increased to 0.3% (0.7% as a whole) in September and October. We conclude that the incidence of aseptic meningitis after MMR vaccination seems to be higher than that reported previously.

Chitosan accelerates the production of osteopontin from polymorphonuclear leukocytes.

Chitosan is a copolymer of beta(1 --> 4) glucosamine and N-acetyl-D-glucosamine, which accelerates the infiltration of polymorphonuclear leukocytes (PMN) in the early phase of wound healing. In the granulation tissue treated with chitosan in canine experimental wound, osteopontin (OPN) was strongly positive in PMN immunohistochemically. OPN is a glycosylated phosphoprotein and promotes the attachment or spread of a variety of cell types. In addition, OPN may play a role in granulomatous inflammation. Production of OPN in PMN was therefore investigated in vitro using human PMN in this study. PMN stimulated with granulocyte-colony stimulating factor (G-CSF) and chitosan accumulated OPN mRNA, and released OPN into their culture supernatants. These findings suggest that OPN is synthesized by migrating PMN which plays the novel role of regulating the evolution of wound healing with chitosan treatment at the early phase of healing.

Effects of chitin and its derivatives on the proliferation and cytokine production of fibroblasts in vitro.

The effects of chitin and its derivatives on the proliferation of fibroblasts and on the production of cytokines were examined in vitro. Chitin and its derivatives showed almost no acceleratory effect on the proliferation of cultured fibroblasts. On the contrary, high-concentration 500 micrograms ml-1) D-glucosamine cultures supplemented with or without a 10% fetal calf serum (FCS) supplementation showed a significant (P < 0.05) reduction in the rate of proliferation of L929 fibroblast cells relative to control. High-concentration chitosan cultures supplemented with 10% FCS showed a significant (P < 0.05) reduction in the rate of L929 fibroblast proliferation. However, the inhibition of cell proliferation by high concentrations of chitosan did not show in cultures without FCS. Interleukin-8 (IL-8) was induced in the supernatants of rat primary cultured dermal fibroblasts stimulated with chitin and its derivatives. Chitin and its derivatives did not stimulate the production of IL-6 by mouse dermal primary cultured fibroblasts. IL-1 alpha, IL-1 beta and tumour necrosis factor-alpha were not detected in the fibroblast supernatants. These observations support the notion that cell proliferation is accelerated indirectly by chitin and its derivatives when these materials are used in vivo. In vivo findings of a angiogenesis and migration of neutrophils may be due to persistent release of IL-8 from fibroblasts.

Accelerating effects of chitosan for healing at early phase of experimental open wound in dogs.

Chitosan is a polymeric beta(1 --> 4) glucosamine (2-amino-2-deoxy-D-glucose) and N-acetyl-D-glucosamine (2-acetamido-2-deoxy-D-glucose) which has been reported as a wound healing accelerator. In order to evaluate the efficacy of chitosan as an accelerator of wound healing, experimental open skin wounds were made on the dorsal side in three normal beagles. Cottonfiber-type chitosan (degree of acetylation = 18%) was applied for 15 days, and the process of wound healing was evaluated histologically and immunohistochemically. On day 3 postwounding, the chitosan-treated wounds showed histologically severe infiltration of polymorphonuclear (PMN) cells and an increase in effusion compared with that in the control. Granulation was more pronounced by the chitosan treatment on day 9 and 15 postwounding. Immunohistochemical typing of collagen I, III and IV showed increase of the production of type III collagen in the chitosan group. The appearance of mitotic cells occurred numerously in the control on postwounding day 3, and in the chitosan group on postwounding day 6. These results suggest chitosan to be having a function in the acceleration of infiltration of PMN cells at the early stage of wound healing, followed by the production of collagen by fibroblasts.

Improvement of the oral bioavailability of digitalis glycosides by cyclodextrin complexation.

Inclusion complexes of the digitalis glycosides digitoxin, digoxin, and methyl digoxin with three cyclodextrins (alpha-, beta-, gamma-homologues) in water and in the solid state were studied by a solubility method, IR and 1H-NMR spectroscopy, and X-ray diffractometry. Solid complexes (in a molar ratio of 1:4) of the digitalis glycosides with gamma-cyclodextrin were prepared and their in vivo absorption examined. The rapidly dissolving form of the gamma-cyclodextrin complex significantly increased plasma levels of digoxin (approximately 5.4-fold) after oral administration to dogs.

Analytical use of solvent extraction with acetonitrile/water/chloroform and 1-propanol/water/cyclohexane mixtures.

To obtain an organic phase containing acetonitrile or 1-propanol from aqueous acetonitrile or aqueous 1-propanol solution, the effect of chloroform and cyclohexane as auxiliary solvents has been investigated. Use of such ternary systems offers an alternative to the so-called salting-out method, in which inorganic salts or hydrophilic organic materials are used to separate organic phases from otherwise miscible solvent mixtures. It also seems preferable for solvent-extraction procedures, since only a small amount of auxiliary solvent is needed instead of the usually large amount of a salt (impurities in which may cause undue contamination); also, the volume and composition of the organic phase can be predicted from phase diagrams and the overall composition of the solvent mixture. Volume-fraction diagrams are especially easy to use. Furthermore, equilibrium is attained in solvent mixtures more rapidly than in salting-out systems. The utility of the ternary solvent systems has been demonstrated for extraction of intermediate molybdophosphate complexes which are specifically formed in aqueous acetonitrile solutions.

The polished precipitate electrode: A new voltammetric method with the solid electrode.

A new surface-renewal technique at the solid electrode has been developed, based on continuous polishing. Well-defined and reproducible current-voltage curves similar to those obtained in polarography are given by the "polished precipitate electrode" (PPE). The method can be used for the continuous determination of electroactive substances and for the study of electrode reaction mechanisms at the solid-liquid interface.

Electrolytic reduction of molybdophosphate in aqueous acetonitrile and its application to flow-coulometric determination of orthophosphate.

The formation and electrolytic reduction of molybdophosphate in aqueous solutions of various water-miscible organic solvents have been extensively investigated. Acetonitrile was found to be the most useful of these solvents. Two species of molybdophosphate are formed in aqueous acetonitrile, one of which changes spontaneously into the other, which is quite stable and undergoes a 2-electron electrolytic reduction. On the basis of these facts, a flow-coulometric method for orthophosphate has been developed, applicable to the range 5 x 10(-6)-1 x 10(-3)M. It has been used for determination of orthophosphate in several phosphorus compounds, some of which are acid-labile.

Determination of metals by solid-liquid separation after liquid-liquid extraction Polarographic determination of indium after extraction with oxine into molten naphthalene.

Indium has been extracted as its oxinate into molten naphthalene, the naphthalene allowed to solidify and then separated and dissolved in dimethylformamide, and the indium determined polarographically in dimethylforiaamide medium with pyridium perchlorate as supporting electrolyte.