Successful Retransplantation With Killer Cell Immunoglobulin-like Receptor Ligand-mismatched Cord Blood in Infant Acute Lymphoblastic Leukemia That Relapsed After Transplantation.

The prognosis of children with KMT2A -rearranged ( KMT2A -r) acute lymphoblastic leukemia (ALL) remains dismal. This report describes the successful retransplantation of a patient with infant ALL who relapsed both bone marrow and central nervous system. The patient received HLA-matched cord blood transplantation (CBT) and relapsed 18 months later. After achieving the second remission, the patient received a killer cell immunoglobulin-like receptor ligand-mismatched CBT with a reduced-intensity conditioning regimen and has been in remission for 52 months. Thus, killer cell immunoglobulin-like receptor ligand-mismatched CBT with reduced-intensity conditioning might be a treatment option for patients with KMT2A- r ALL who relapsed after transplantation, even with extramedullary relapse.

Pelvic Tilt Reduces the Accuracy of Acetabular Component Placement When Using a Portable Navigation System: An In Vitro Study.

BACKGROUND: A portable navigation system (PNS) was recently introduced. The PNS enables surgeons to place the acetabular component accurately. While the margin of the error for the cup abduction and anteversion was larger than the values obtained from a computed tomography-based navigation system. We hypothesized that the accuracy of the PNS might be affected by pelvic tilt. MATERIAL AND METHODS: A bone substitute model of the pelvis was used in this in vitro study. We set the acetabular component using PNS. We set the acetabular component angle after changing the sagittal, coronal, and axial pelvic tilt. We calculated the difference between the angle displayed on the PNS display and the actual angle of the acetabular component. The difference in inclination angle was defined as ΔRI, and the difference in the anteversion angle was defined as ΔRA. We evaluated the trends in this ΔRI and ΔRA due to the pelvic tilt. RESULTS: In this in vitro study, the placement of the acetabular component was accurate in the neutral position; ΔRI was 0.5 ± 0.7° and ΔRA was 1.0 ± 0.7°. Sagittal pelvic tilt and axial pelvic tilt increased both the ΔRA and ΔRI (P = .017). Coronal tilt increased ΔRI but did not change ΔRA. CONCLUSIONS: While the PNS may enable surgeons to place accurate component placement in the neutral position, its accuracy decreased by pelvic tilt. The surgeons should use a solid pelvic lateral positioner for reducing discrepancies in pelvic tilt when using the PNS in the lateral decubitus position.

Recurrent Shoulder Tip Pain After Ventriculoperitoneal Shunt Placement Associated with Infectious Peritonitis with Propionibacterium acnes; A Case Report and Review of the Literature.

Ventriculoperitoneal (VP) shunt placement is commonly performed for the treatment of hydrocephalus, and several complications of this procedure are well known. Radiating shoulder tip pain after VP shunt placement has been reported as an unusual complication in a few cases, associated with dislocation of the peritoneal catheter. We described the case of a 9-year-old girl who presented with recurrent radiating shoulder tip pain after VP shunt placement. The pain recurred after peritoneal catheter repositioning because of peritoneal inflammation and adhesion due to peritonitis with Propionibacterium acnes (P. acnes). This bacterium was isolated using 16S ribosomal RNA gene polymerase chain reaction (16S rRNA gene PCR), and anaerobic and prolonged culture tests. After antibacterial treatment, ventriculoarterial (VA) shunt placement was successfully performed. Hemidiaphragm irritation by the peritoneal catheter leads to radiating shoulder tip pain, and peritoneal inflammation and adhesion caused by infectious peritonitis may cause recurrence of this despite catheter repositioning. Clinicians should be aware of shoulder pain as a complication of VP shunt placement, and should consider VA shunt placement as an alternative treatment if this symptom recurs after catheter repositioning. Furthermore, 16S rRNA gene PCR and anaerobic and prolonged culture tests should be considered to detect P. acnes infection.

[A Pediatric Case of Xanthogranuloma in the Suprasellar Region Detected by a Severe Short Stature after 6 Years Growth Failure].

Here we report a case of a 12-year-old girl who was referred to our department because of marked short stature of more than -5 SD below the median. Although her growth failure began suddenly at 6 years of age, she never had an examination because she had no other symptoms. Brain MRI examination suggested a tumor in the suprasellar region, and endocrine examination revealed combined pituitery hormone deficiency due to the tumor. Before surgery, the supplementation with hydrocortisone and levothyroxine was initiated. The pathological diagnosis of the surgically removed tumor was xanthogranuloma. The pattern of her growth curve showed a growth failure with sudden onset, which is a typical pattern of short stature secondary to pituitary disfunction including growth hormone deficiency associated with brain tumors. This case suggests that growth failure could be the only symptom in pediatric cases with brain tumors. Improved awareness regarding the association of growth failure with brain tumors is needed for earlier diagnosis and treatment. Furthermore, the growth curves should be carefully evaluated in regular health examinations at school.

[A Pediatric Case of Acquired Hemophilia A: The Usefulness of the Activated Partial Thromboplastin Time (APTT) Cross-Mixing Test for Early Diagnosis].

Acquired hemophilia A (AHA), a bleeding disorder caused by autoantibodies against FVIII, has the potential for life-threatening bleeding. The annual onset rate is said to be one in 4 million people, but diagnosis examples increase in adults because a disorder concept penetrated. AHA is quite rare in children, with an incidence rate of 0.045 per 1 million, but early detection is crucial because serious bleeding can happen, as in adults. We report a pediatric case who received an early diagnosis of AHA by an activated partial thromboplastin time (APTT) cross-mixing test. The 12-year-old girl had neither a past history nor a family history of bleeding episodes. She presented with intramuscular bleeding and epistaxis without trauma or medication. At diagnosis, her blood test showed prolonged APTT. Other hemostatic tests, such as the platelet count, prothrombin time and fibrinogen concentration, were within the normal range. We administered an APTT cross-mixing test that detected an inhibitor pattern and inhibitory antibodies against factors VIII. As a result, we administered prednisolone and the inhibitor disappeared after 1.5 months. In conclusion, AHA is a bleeding disorder which should be considered even in children due to the potential for life-threatening bleeding. Furthermore, the APTT cross-mixing test is useful for screening coagulation factor deficiencies and inhibitors.

Contraction and extension of Vorticella and its mechanical characterization under flow loading.

We have studied the contraction and extension of Vorticella convallaria and its mechanical properties with a microfluidic loading system. Cells of V. convallaria were injected to a microfluidic channel (500 µm in width and 100 µm in height) and loaded by flow up to ∼350 mm s(-1). The flow produced a drag force on the order of nanonewton on a typical vorticellid cell body. We gradually increased the loading force on the same V. convallaria specimen and examined its mechanical property and stalk motion of V. convallaria. With greater drag forces, the contraction distance linearly decreased; the contracted length was close to around 90% of the stretched length. We estimated the drag force on Vorticella in the channel by calculating the force on a sphere in a linear shear flow.

Three-dimensional two-component velocity measurement of the flow field induced by the Vorticella picta microorganism using a confocal microparticle image velocimetry technique.

Understanding the biological feeding strategy and characteristics of a microorganism as an actuator requires the detailed and quantitative measurement of flow velocity and flow rate induced by the microorganism. Although some velocimetry methods have been applied to examine the flow, the measured dimensions were limited to at most two-dimensional two-component measurements. Here we have developed a method to measure three-dimensional two-component flow velocity fields generated by the microorganism Vorticella picta using a piezoscanner and a confocal microscope. We obtained the two-component velocities of the flow field in a two-dimensional plane denoted as the XY plane, with an observation area of 455x341 mum(2) and the resolution of 9.09 mum per each velocity vector by a confocal microparticle image velocimetry technique. The measurement of the flow field at each height took 37.5 ms, and it was repeated in 16 planes with a 2.50 mum separation in the Z direction. We reconstructed the three-dimensional two-component flow velocity field. From the reconstructed data, the flow velocity field [u((x,y,z)),v((x,y,z))] in an arbitrary plane can be visualized. The flow rates through YZ and ZX planes were also calculated. During feeding, we examined a suction flow to the mouth of the Vorticella picta and measured it to be to 300 pls.

Stereoselective synthesis of beta-hydroxyphenylalanines using imino 1,2-Wittig rearrangement of hydroximates.

The imino 1,2-Wittig rearrangement of hydroximates containing a furan ring provides a novel method for the synthesis of beta-hydroxy-alpha-amino acids. Upon treatment with LDA, hydroximates smoothly underwent the rearrangement to give Z-2-hydroxyoxime ethers in good yield, which were converted into both cis- and trans-oxazolidinones with high stereoselectivity. The cis- and trans-oxazolidinones were stereoselectively converted into erythro- and threo-beta-hydroxyphenylalanines, respectively, via the oxidative cleavage of a furan ring, ring-opening of oxazolidinone, and deprotection.

Chemical modification of glycerinated stalks shows tyrosine residues essential for spasmoneme contraction of Vorticella sp.

Chemical modification of glycerinated stalks of Vorticella with TNM is used to investigate the role of tyrosine residues in the Ca(2+)-induced contraction of the spasmoneme. Tetranitromethane (TNM) is often employed as a specific reagent for the nitration of tyrosine residues in a protein at neutral and slightly alkaline pHs although TNM can also oxidize cysteine residues in the acidic and neutral pH range. Prior incubation with Ca(2+) of stalks to be treated with TNM can protect the spasmoneme from irreversible denaturation. On the other hand, TNM treatment in the absence of free Ca(2+) causes an irreversible denaturation of the spasmoneme. It was revealed by us that an isolated Ca(2+)-binding protein called spasmin could not bind with Ca(2+) after TNM treatment, even if the treatment was performed in the presence of Ca(2+). In an additional experiment, we confirmed that the chemical modification of cysteine residues in the spasmoneme with N-7-dimethyl-amino-4methyl- coumarinyl- maleimide (DACM) has no effect on the contractibility. These results suggest that tyrosine residues in spasmin are essential for spasmoneme contraction and are protected from TNM in the presence of Ca(2+) when spasmin binds with its receptor protein in the spasmoneme.

Measurement of circular birefringence and circular dichroism of the single crystals of lambda-(+)589- and delta-(-)589-tris(ethylenediamine)cobalt(III) triiodide monohydrate by the extended HAUP method.

We have achieved measuring four optical parameters simultaneously, namely, linear birefringence (LB), circular birefringence (CB), linear dichroism (LD), and circular dichroism (CD), of single crystals of Lambda-(+)(589)- and Delta-(-)(589)-tris(ethylenediamine) cobalt(III) triiodide monohydrate (1) along the <001> plane at the fixed wavelength (514.5 nm). Such measurements are possible only when the High Accuracy Universal Polarimeter (HAUP) is employed; it is called the extended HAUP method. Our experimental results showed that both LB and LD of the Lambda-(+)(589)-(1) crystal have the same magnitude as those of the Delta-(-)(589)-(1) crystal. It was also revealed for the first time that the CB data of crystals of Lambda-(+)(589)-(1) and Delta-(-)(589)-(1) are almost of the same magnitude, but are of opposite sign, reflecting their opposite absolute configurations. On the other hand, although the CD data obtained for Lambda-(+)(589)-(1) is almost three times larger than that for Delta-(-)(589)-(1,) these CD data are also opposite in sign, as expected from the opposite chirality of crystals. .

Detection of neutrophils in late-onset interface inflammation associated with flap injury after laser in situ keratomileusis.

OBJECTIVE: To report a case with late-onset interface inflammation associated with traumatic flap injury at 7 months after laser in situ keratomileusis (LASIK) and to describe the type of infiltrating cells in the tears of the patient. METHODS: Interventional case report. A 24-year-old male patient who underwent uneventful LASIK on both eyes received blunt trauma from the tip of a shoe in the left eye 7 months after surgery. The corneal flap of his left eye was lacerated across the pupillary area. Inflammatory cells were observed under the lacerated flap segment. Tear fluid was collected from his left eye 3 days after the injury and assessed by tear cytology. For controls, tears of 2 patients who underwent LASIK and developed no interface inflammation were collected the next day after their surgeries and examined. RESULTS: Tear fluid of the patient with interface inflammation contained numerous neutrophils. Tears of control patients contained only a few epithelial cells and cell debris but no inflammatory cells. The inflammation was decreased by systemic and topical steroids. However, irregular astigmatism caused by stromal scarring remained, resulting in decreased best-corrected visual acuity. CONCLUSIONS: Interface inflammation can be caused by late-onset flap injury. Neutrophils detected in the tears may reflect a major component of cells infiltrating the interface after LASIK.

Novel nuclear and cytoplasmic proteins detected by anti-Zoothamnium arbuscula (Protozoa) spasmin 1 antibody in mammalian cells are dependent on the cell cycle.

Spasmin is a calcium-binding protein that is the major component of calcium-induced contractile filaments, called spasmoneme, found in vorticellid ciliates. Such filaments have not been observed in any organisms other than green algae. To determine whether calcium-induced contractile filaments resembling spasmoneme are present in higher eukaryotes, we performed immunofluorescence imaging of an anti-Zoothamnium arbuscula (protozoa, ciliophora) spasmin 1 polyclonal antibody in HeLa cells. In the cytoplasm, ubiquitous antigens seemed to be co-localized with microtubules at interphase, but not throughout mitosis. In the nucleus, areas linked to the nuclear envelope contained a number of hot spots. These regions were unclear during condensation of the replicated chromosomes, but became clearly visible again at cytokinesis. Immunoblotting analysis identified localized antigens during different phases of the cell cycle, including a 68/71 kDa cytoplasmic protein and a 55 kDa nuclear protein in interphase, and a 55/70 kDa protein in mitosis. The anti-spasmin 1 antibody recognized antigens in both hamster kidney BHK21 cells and Human lung cancer A-549 cells. These results suggest that novel spasmin-like proteins could be common in mammalian cells.

Chemical modification of contractile 3-nm-diameter filaments in Vorticella spasmoneme by diethyl-pyrocarbonate and its reversible renaturation by hydroxylamine.

A peritrich ciliate possesses a zooid and a long stalk consisting of a bundle of 3-nm-diameter filaments. Glycerinated stalks can contract in the presence of free Ca(2+) and re-extend in the absence of free Ca(2+). In the present study, we demonstrated that histidine residue(s) played a critical role in spasmoneme contraction by using glycerinated stalk of Vorticella. Concentration-dependent inhibition of spasmoneme contraction was observed in the presence of reversible histidine-modifying reagent named diethyl-pyrocarbonate (DEPC). In addition, the contractility degree of DEPC-modified spasmoneme could be partially restored by hydroxylamine treatment. The 244nm absorption of modified spasmoneme protein(s) increased with rising DEPC concentration and decreased following the addition of hydroxylamine treatment. Adding Ca(2+) before DEPC modification could prevent the spasmoneme contraction from inhibition of DEPC. Those results suggested that histidine residues were actively involved in spasmoneme contraction. Ca(2+)-binding ability of spasmin was not inhibited by DEPC modification, which suggested that the essential histidine residues were not on the calcium-binding site of spasmin.

Characterization of the spasmin 1 gene in Zoothamnium arbuscula strain Kawagoe (protozoa, ciliophora) and its relation to other spasmins and centrins.

Zoothamnium arbuscula strain Kawagoe is a giant sessile peritrich ciliated protozoa that possesses a contractile organelle called a spasmoneme. We report here on the molecular characterization and provide an opportunity to discuss the evolutionary relationships of the Z. arbuscula spasmin; spasmins belong to the calmodulin superfamily and are the major components of spasmoneme filaments. We analysed and obtained the whole sequence of the spasmin 1 gene and a partial sequence of the spasmin 2 gene. It is surprising that the sequence of spasmin 1 does not contain introns and encodes an open reading frame of 531 bp. It predicts a product of 177 amino acids with a calculated molecular mass of 19659 Da and a pI of 4.68. The amino acid sequence has two putative calcium-binding domains. One of them is a functional domain, as defined by the EF-hand consensus. The varieties of spasmins were revealed by comparison with amino acid components and molecular relationships of spasmin 1 protein and other spasmins. A comparison of the amino acid sequence between the Z. arbuscula spasmin and known centrins indicates that spasmins have a one residue deletion in the EF-hand domain-2 and four residue insertions in domain-4, as does the Vorticella spasmin. However, there are large variations in the amino acid sequence at domain-4 within spasmin 1, spasmin 2 and the Vorticella spasmin.

Phylogenetic relationships between Vorticella convallaria and other species inferred from small subunit rRNA gene sequences.

Vorticellid ciliates generally dwell in freshwater. In nature, the species have up until now been identified by comparison with previous descriptions. It is difficult to identify between species of the genus Vorticella, because the morphological markers of vorticellid ciliates described in reports are limited and variable. Unfortunately, culturing them has only succeeded with certain species such as Vorticella convallaria, but many others have been impossible to culture. To find out whether the sequence of a small subunit rRNA gene was an appropriate marker to identify vorticellid ciliates, the gene was aligned and compared. Finding a new convenient method will contribute to research on vorticellid ciliates. In strains of V. convallaria, classified morphologically, some varieties of the SSrRNA gene sequences were recognized, but there were large variations within the same species. According to the phylogenetic tree, these strains are closely related. However, the difference was not as big as between Vorticella and Carchesium. In addition, Carchesium constructed a distinct clade from the genus Vorticella and Epistylis. These results show the possibility that the SSrRNA gene is one of the important markers to identify species of Vorticella. This study is first to approach and clarify the complicated taxa in the genus Vorticella.