The correlation of urinary podocytes and podocalyxin with histological features of lupus nephritis.

Objectives The objective of this study was to test the correlation of urinary podocyte number (U-Pod) and urinary podocalyxin levels (U-PCX) with histology of lupus nephritis. Methods This was an observational, cross-sectional study. Sixty-four patients were enrolled: 40 with lupus nephritis and 24 without lupus nephritis (12 lupus nephritis patients in complete remission and 12 systemic lupus erythematosus patients without lupus nephritis). Urine samples were collected before initiating treatment. U-Pod was determined by counting podocalyxin-positive cells, and U-PCX was measured by sandwich ELISA, normalized to urinary creatinine levels (U-Pod/Cr, U-PCX/Cr). Results Lupus nephritis patients showed significantly higher U-Pod/Cr and U-PCX/Cr compared with patients without lupus nephritis. U-Pod/Cr was high in proliferative lupus nephritis (class III±V/IV±V), especially in pure class IV (4.57 (2.02-16.75)), but low in pure class V (0.30 (0.00-0.71)). U-Pod/Cr showed a positive correlation with activity index ( r=0.50, P=0.0012) and was independently associated with cellular crescent formation. In contrast, U-PCX/Cr was high in both proliferative and membranous lupus nephritis. Receiver operating characteristic analysis revealed significant correlation of U-Pod/Cr with pure class IV, class IV±V and cellular crescent formation, and the combined values of U-Pod/Cr and U-PCX/Cr were shown to be associated with pure class V. Conclusions U-Pod/Cr and U-PCX/Cr correlate with histological features of lupus nephritis.

Impact of the COVID-19 pandemic on the incidence of surgical site infection after orthopaedic surgery: an interrupted time series analysis of the nationwide surveillance database in Japan.

BACKGROUND: During the COVID-19 pandemic, hygiene awareness was increased in communities and hospitals. However, there is controversy regarding whether such circumstances affected the incidence of surgical site infections (SSIs) in the orthopaedic surgical field. AIM: To examine the impact of the COVID-19 pandemic on the incidence of SSIs after orthopaedic surgery. METHODS: The medical records of patients having undergone orthopaedic surgery were extracted from the nationwide surveillance database in Japan. The primary outcomes were the monthly incidences of total SSIs, deep or organ/space SSIs, and SSIs due to meticillin-resistant Staphylococcus aureus (MRSA). Interrupted time series analysis was conducted between pre-pandemic (January 2017 to March 2020) and pandemic (April 2020 to June 2021) periods. RESULTS: A total of 309,341 operations were included. Interrupted time series analysis adjusted for seasonality showed no significant changes in the incidence of total SSIs (rate ratio 0.94 and 95% confidence interval 0.98-1.02), deep or organ/space SSIs (0.91, 0.72-1.15), or SSIs due to MRSA (1.07, 0.68-1.68) along with no remarkable slope changes in any parameter (1.00, 0.98-1.02; 1.00, 0.97-1.02; and 0.98, 0.93-1.03, respectively). CONCLUSIONS: Awareness and measures against the COVID-19 pandemic did not markedly influence the incidence of total SSIs, deep or organ/space SSIs, or SSIs due to MRSA following orthopaedic surgery in Japan.

Validation study of the Japanese version of the Faecal Incontinence Quality of Life Scale.

AIM: The aim of the present study was to conduct a psychometric validation of the Japanese version of the FIQL (JFIQL). METHOD: A retrospective analysis of data from the JFIQL was conducted. Wexner scores and Faecal Incontinence Severity Index (FISI) scores were collected prospectively in patients with faecal incontinence who visited our centre between 2008 and 2009. For convergent validity, the JFIQL scores were compared with stages on the Wexner scale for lifestyle alteration. To evaluate reliability, Cronbach's alpha was calculated for internal consistency, whereas a test-retest study was performed to evaluate reproducibility. In assessing responsiveness, JFIQL scores before and after treatments were compared in patients whose FISI scores decreased by ≥ 50%. RESULTS: Convergent validity and internal consistency were determined in 70 patients (49 women; median age 68.5 years). The JFIQL scores were significantly associated with lifestyle alteration stages on the Wexner scale, demonstrating convergent validity in all four domains and the generic score. Cronbach's alpha was > 0.7 for generic scores and all domains except Embarrassment. The intraclass correlations for the 27 patients available for the test-retest study were > 0.7 for generic scores and all domains except Embarrassment. The median JFIQL score improved significantly after treatment in the 23 patients whose FISI scores decreased ≥ 50%, indicating good responsiveness in all four domains and the generic score. CONCLUSION: The JFIQL has been validated and is now ready for use in evaluating the symptom-specific quality of life in Japanese patients with faecal incontinence.

Autoantibodies specific for different isoforms of CD45 in systemic lupus erythematosus.

Nearly one-third of IgM antilymphocyte autoantibody-positive sera from patients with systemic lupus erythematosus (SLE) contain IgM antibodies to one or more 180-220-kD molecules (p180, p190, p205, and p220) in blots of glycoproteins purified from T cells by wheat germ agglutinin affinity chromatography. Identity of these IgM targets with multiple isoforms of CD45 was established by their specific depletion from T cell glycoproteins by immunoprecipitation with T191, a monoclonal antibody (mAb) that reacts with an epitope common to all CD45 isoforms. Although the anti-CD45 autoantibodies recognize higher molecular weight isoforms primarily, antigenic specificity in this system is quite heterogeneous and includes multiple distinct CD45 isoforms on different types of T cells that are, at least in part, different from those reactive with mAbs 2H4 and UCHL-1. Because CD45 is a major membrane protein tyrosine phosphatase that plays a critical role in antigen-induced T cell activation, the present data may be relevant to some of the antilymphocyte antibody-mediated immunologic abnormalities that characterize SLE and related autoimmune diseases.

Constitutive expression of a groEL-related protein on the surface of human gamma/delta cells.

Rabbit antibodies to hsp58 (P1), the human homologue of the Escherichia coli stress protein groEL, react specifically in indirect immunofluorescence and complement-dependent microcytoxicity experiments with a cell surface antigen expressed constitutively by T cell lines bearing gamma/delta receptors. This anti-hsp58-reactive antigen is not demonstrable on T cells that express alpha/beta receptors or on various cells that lack T cell receptors. Certain evidence was obtained to suggest that the target antigen on the surface of gamma/delta T cells is a approximately 77-kD protein distinct from intracellular hsp58 and known members of the hsp70 stress protein family. While the exact nature and significance of this anti-hsp58-reactive protein remain to be determined, these data may help to clarify the roles of groEL-related stress proteins and gamma/delta cells that recognize groEL homologous in immunologic defense against infection and in autoimmune disease.

Detection of umbilical venous constriction by Doppler flow measurement at midgestation.

OBJECTIVES: To investigate whether umbilical venous velocity and venous velocity pulsation are associated with umbilical vein diameter, umbilical ring diameter and umbilical cord coiling index at midgestation. METHODS: Two hundred and eighty pregnant women were enrolled in the study at between 18 and 24 weeks of gestation. The diameter of the umbilical cord and internal diameter of the umbilical vein in a free loop and at the ring, and the umbilical coiling index, were measured using ultrasonography. Umbilical venous velocities were measured by Doppler ultrasonography at the umbilical ring and a free loop of the cord. RESULTS: All variables were successfully measured in 92% of the patients. There were negative correlations between the diameters of the umbilical ring and of the umbilical vein at the ring and the venous velocity at the umbilical ring. The venous velocity at the umbilical ring was significantly higher and the umbilical ring diameter was significantly lower in fetuses with umbilical venous pulsation at the free loop. Significant correlations were observed between the venous velocity and amplitude of pulsation. Venous pulsations at the free loop were frequently observed in fetuses with a hypercoiled cord. CONCLUSION: High venous velocity and increased venous pulsation at the umbilical ring may be associated with umbilical cord constriction.

Predisposing factors for massive hemorrhage during Cesarean section in patients with placenta previa.

OBJECTIVES: To investigate whether maternal history and ultrasound findings can be predictors for massive hemorrhage during Cesarean section in patients with placenta previa and adherence of the placenta. METHODS: We reviewed 127 singleton pregnancies with placenta previa. Maternal history, antenatal ultrasound findings of the placenta, including location, presence of placental lacunae, lack of a clear zone, presence of sponge-like findings of the cervix and presence of a marginal sinus in cases of placenta previa were reviewed retrospectively, and their association with amount of bleeding during Cesarean section was analyzed. RESULTS: Logistic regression analysis revealed that advanced maternal age (odds ratio (OR), 5.4; 95% CI, 1.8-16.4), previous Cesarean section (OR, 20.4; 95% CI, 4.0-105.2) and sponge-like findings in the cervix (OR, 5.6; 95% CI, 1.8-17.0) were associated with massive bleeding (> 2500 mL). Placental adherence occurred in five cases and was more frequent in cases where the placenta was located at the site of the scar of a previous Cesarean section (OR, 123.1; 95% CI, 4.5-3395.2) and where there was lack of a clear zone (OR, 48.0; 95% CI, 3.8-604.7). CONCLUSIONS: Advanced maternal age, previous Cesarean section and presence of sponge-like findings in the cervix are risk factors for massive bleeding during Cesarean section in cases of placenta previa, regardless of whether placental adherence is present. Placental location on the scar of a previous Cesarean section and lack of a clear zone are risk factors for placental adherence. When these findings are identified preoperatively, management should be tailored accordingly.

A novel exogenous concentration-gradient collagen scaffold augments full-thickness articular cartilage repair.

OBJECTIVES: A collagen scaffold has been long used in order to enhance the regeneration of articular cartilage. In the present study, we investigate the effectiveness of a concentration-gradient (CG) collagen that is designed to recruit efficiently the mesenchymal stem cells (MSCs) to the central region of the full-thickness cartilage defects via haptotaxis. METHODS: The present study used Cellmatrix (0.3% type I collagen; Nitta gelatin, Osaka, Japan) as the collagen material. We prepared 33%CG collagen gel and 50%CG collagen gel. No gradient collagen gel served as negative control. Full-thickness cartilage defects were created at the patella groove of the rabbit knee, to which the three different collagen gels were transplanted. Bromodeoxyuridine (BrdU) positive, proliferating cells were enumerated and localized, whereas the histological grading score for cartilage regeneration was counted. The expression of type I and type II collagens was evaluated by immunohistochemistry. We also confirmed that the MSCs migrate toward the collagen substrate of higher concentration in a stringently in vitro haptotactic manner. RESULTS: Enumeration of the BrdU-positive cells demonstrated that 33%CG collagen gel recruited a significantly larger number of proliferating cells to the central region of the cartilage defect. The histological grading score for the regenerated cartilage treated with 33%CG collagen gel was superior to the other groups. CONCLUSIONS: CG collagen scaffold recruits effectively the MSCs to the center of full-thickness cartilage defect and enhances regeneration of the full-thickness cartilage defect.

Vascular compliance is secured under angiotensin inhibition in non-diabetic chronic kidney diseases.

Cardiovascular diseases constitute major cause of death in chronic kidney diseases (CKDs). We examined the effects of angiotensin inhibition either with angiotensin-converting enzyme inhibitor or with angiotensin receptor blocker on patient prognosis and heart-ankle pulse wave velocity (haPWV) in CKDs. Randomized controlled study was performed on 102 patients with non-diabetic CKDs. Patients were divided into two groups with or without angiotensin inhibition, and followed until death, creatinine clearance was halved or starting renal replacement therapy, whichever occurred first. For 4 years, haPWV was assessed repeatedly in the surviving patients. While both groups showed well blood pressure control throughout 4 years (129+/-1 to 131+/-2/71+/-1 to 73+/-2 mm Hg), renal prognosis was better in angiotensin inhibition group (P<0.05). In addition, angiotensin inhibition reduced cardiovascular and renal death (P<0.05). Age, sex, heart rate, systolic blood pressure and proteinuria were correlated to haPWV (R(2)=0.76, P<0.0001). Although haPWV was similar between two groups at the start of the study (1098+/-31 vs 1094+/-37 cm/s), it was higher in patients without angiotensin inhibition than that with angiotensin inhibition 4 years later (1034+/-38 cm/s (n=28) vs 1242+/-37 cm/s (n=23), P<0.01). The present results provided the evidence that angiotensin inhibition arrested a time-dependent elevation of haPWV in non-diabetic CKDs, conferring organ protection. Furthermore, our data indicated that angiotensin inhibition improved patient prognosis in non-diabetic chronic kidney diseases with mild-to-moderate renal dysfunction.

Changes of vascular endothelial growth factor after vitrectomy for macular edema secondary to retinal vein occlusion.

PURPOSE: To examine whether vitrectomy combined with retinal photocoagulation reduces the vitreous level of vascular endothelial growth factor (VEGF) in patients with macular edema associated with retinal vein occlusion (RVO). METHODS: The authors measured VEGF levels in vitreous samples from four eyes of four patients with RVO during vitrectomy and fluid samples obtained during revitrectomy 3 to 9 months postoperatively for complications: an epiretinal membrane in two patients, macular holes in one patient, and vitreous hemorrhage in one patient. During vitrectomy, retinal photocoagulation was performed on the ischemic region of the retina in all cases (mean of 510 shots). RESULTS: In four eyes with RVO, there was a difference in the vitreous VEGF levels between the vitreous samples obtained during vitrectomy (mean of 2692 pg/mL, range of 15.6-9040 pg/mL) and the fluid samples obtained at the time of revitrectomy (mean of 947 pg/mL, range of 15.6-3430 pg/mL). CONCLUSIONS: The results suggest that the vitreous levels of VEGF may be reduced by vitrectomy combined with retinal photocoagulation for macular edema with RVO. It may be important to reduce the vitreous levels of VEGF by vitrectomy and retinal photocoagulation for ischemic retina in macular edema with RVO.

Intratumoural expression of thymidylate synthase is an independent predictor of prognosis in patients with squamous cell carcinoma of the tongue: results from a retrospective study.

The aim of this study was to assess the importance of immunohistochemical thymidylate synthase (TS) expression level as a prognostic marker in tongue cancer patients. In 140 patients with primary squamous cell carcinoma (SCC) of the tongue, intratumoural TS expression was evaluated by immunohistochemistry. The level of TS expression was determined by a semiquantitative scoring system, ranging from 1+ to 3+ according to the ratio of TS-positive cells. Of 140 patients, 64 (45.7%), 49 (35.0%) and 27 (19.3%) were assessed as 1+, 2+ and 3+, respectively. Univariate analyses demonstrated that both disease-free survival (DFS) and overall survival (OS) were significantly lower in patients with a TS 3+ tumour than in those with a TS 1+/2+ tumour (DFS: P = 0.0082, OS: P = 0.0100). In a multivariate analysis using the Cox regression model, cervical lymph-node status and TS expression level were selected as independent factors for DFS and OS. Maintenance adjuvant chemotherapy by oral 5-fluorouracil (5-FU) significantly improved DFS and OS in patients with a TS 1+/2+ tumour (DFS: P = 0.0027, OS: P = 0.0398). These data suggest that the level of immunohistochemical TS expression is an independent prognosticator in patients with tongue SCC, and may be useful in the selection of patients who would benefit from oral 5-FU adjuvant chemotherapy.

Closed reduction for traumatic posterior dislocation of the shoulder using the 'lever principle': two case reports and a review of the literature.

Traumatic posterior dislocation of the shoulder is frequently missed because of its rarity and the absence of characteristic symptoms. Several signs should be emphasised: an overlap of the humeral head and glenoid rim in a true anteroposterior view and the light-bulb sign in the anteroposterior view. To make an accurate and early diagnosis, use of multidirectional radiographs combined with computed tomography is recommended. Closed reduction was successfully performed under general anaesthesia using the DePalma method with slight modification--the lever principle--by pushing the medial side of the upper arm laterally to adduct the shoulder as far as possible. The dynamics of the lever principle make it a safer and more effective method of achieving a closed reduction of a posterior dislocation of the shoulder than the conventional method of solely pushing the humeral head anteriorly, especially in patients with locking of the glenohumeral joint and impression fractures.

Modulation of the vacuolar H+-ATPase by adenylates as basis for the transient CO2-dependent acidification of the leaf vacuole upon illumination.

Using tonoplast vesicles, we have investigated the activity of the vacuolar H+-ATPase which is the dominant proton pump at the tonoplast of mesophyll cells. Bafilomycin-sensitive ATP hydrolysis or acidification of tonoplast vesicles in the presence of ATP were measured at varying ATP, ADP and Pi concentrations, and in the presence of oxidized or reduced glutathione. Increased ATP/ADP ratios as reported for the extrachloroplast cytoplasm during the induction phase of photosynthesis at high or low CO2 (P. Gardeström, Biochim. Biophys. Acta 1183 (1993) 327-332) increased the activity of the V-ATPase in simulation experiments with vesicles. Depending on reported subsequent decreases in cytoplasmic ATP/ADP ratios in the presence of high or low CO2, the ATPase activity of tonoplast vesicles changed in simulation experiments to lower values. More than 10 mM phosphate was required to decrease the ATPase activity in vesicles significantly at ATP/ADP ratios of 3 or higher, indicating that ATPase activity is controlled more by ratios of ATP to ADP than by phosphorylation potentials (ATP)/(ADP)(Pi). Oxidized glutathione was inhibitory. The results permit interpretation of the observation that on illumination of previously darkened leaves the pH of the vacuoles of mesophyll cells decreases indicating energized transport of protons across the tonoplast into acidic vacuoles, and that the extent of vacuolar acidification depends on the CO2 concentration of the surrounding air (Z.-H. Yin, S. Neimanis, U. Heber, Planta 182 (1990) 253-261). We conclude that short term control of tonoplast ATPase activity in leaves during dark/light transients can essentially be understood on the basis of reported changes in cytoplasmic ATP/ADP ratios, with a possible participation of redox modulation.

cDNA cloning of mouse ferredoxin reductase from kidney.

A cDNA encoding ferredoxin reductase has been isolated from a mouse kidney cDNA library using human ferredoxin reductase cDNA as a probe. Mouse ferredoxin reductase coded for 494 amino acid residues. The mouse mature enzyme which comprises 460 amino acid residues shared 87.8-89.1% amino acid identities with the bovine and human enzyme. Northern blot analysis showed that ferredoxin reductase mRNA was expressed in the adrenal, testis and ovary and to a lesser extent in the liver and kidney. However, this mRNA in the adrenal cell line, Y-1 cell, was not induced by adenosine 3',5'-cyclic monophosphate (cAMP) in contrast with ferredoxin mRNA.

Molecular cloning of 25-hydroxyvitamin D-3 24-hydroxylase (Cyp-24) from mouse kidney: its inducibility by vitamin D-3.

A cDNA encoding a 25-hydroxyvitamin D-3 24-hydroxylase, Cyp-24, has been isolated from mouse kidney cDNA library by hybridization screening. Mouse Cyp-24, coding for 514 amino acid residues, shared 82.1 and 94.7% amino acid identity with human and rat CYP24s, respectively. Among mouse organs examined, Cyp-24 mRNA could be detected in the kidney. When mice were treated with vitamin D-3, Cyp-24 mRNA was induced in the kidney.

Mouse cytochrome P-450 linked ferredoxin: its cDNA cloning and inducibility by dibutyryladenosine 3',5'-cyclic monophosphate and forskolin.

Two full-length cDNAs (F1-1 and F41-1) complementary to mouse kidney mRNA coding for cytochrome P-450 (P450) linked ferredoxin were isolated and completely sequenced. The coding sequences between F1-1 and F41-1 were identical. However, the 3' untranslated regions of F1-1 and F41-1 were 228 and 27 bases long due to the presence of alternative polyadenylation sites, respectively. The deduced amino acid sequence of mouse cytochrome P-450 linked ferredoxin showed 92.5, 75.0, 71.2 and 71.0% identities with those of rat, human, pig and bovine cytochrome P-450 linked ferredoxin, respectively. The cytochrome P-450 linked ferredoxin mRNA was detected in adrenal, kidney and ovary among the organs examined. The treatment of Y-1 cells with dibutyryladenosine 3',5'-cyclic monophosphate or forskolin induced the transcript of cytochrome P-450 linked ferredoxin mRNA.

Molecular cloning and sequencing of the cDNA for vacuolar H+-pyrophosphatase from Chara corallina1.

We have cloned a cDNA for vacuolar proton-translocating pyrophosphatase of Chara corallina that is one of the closest green algae to the land plants. The deduced protein consists of 793 amino acid residues. Its sequence is 71% identical to the H+-pyrophosphatases of land plants, and is less than 46% identical to those of marine alga and phototrophic bacterium.

Simultaneous expression of ferredoxin, ferredoxin reductase and P450 in COS7 cells.

cDNA fragments encoding mouse ferredoxin and ferredoxin reductase were simultaneously introduced into COS7 cells by using an expression vector, pUC-SR alpha plasmid. When using the mitochondrial fraction prepared from the transfected cells, cytochrome-c reductase activity was detected. This activity was highest when 7.5 micrograms of the ferredoxin expression plasmid (pSR alpha F) and 2.5 micrograms of the ferredoxin reductase expression plasmid (pSR alpha FR) were transfected into COS7 cells. In this system, NADPH could be replaced by NADH as a cofactor for the reduction of cytochrome-c although the cytochrome-c reductase was more dependent on NADPH than NADH at a low concentration. When CYP24 expression plasmid was transfected into COS7 cells along with both pSR alpha F and pSR alpha FR, the transfected cells revealed a 3-fold higher 25-hydroxyvitamin D3-24-hydroxylase activity than COS7 cells transfected with CYP24 expression plasmid.

Lack of Control in Inorganic Phosphate Uptake by Catharanthus roseus (L.) G. Don Cells (Cytoplasmic Inorganic Phosphate Homeostasis Depends on the Tonoplast Inorganic Phosphate Transport System?).

Inorganic phosphate (Pi) uptake by Catharanthus roseus (L.) G. Don cells was studied in relation to its apparent uncontrolled uptake using 31P-nuclear magnetic resonance spectroscopy. Kinetics of Pi uptake by the cells indicated that apparent Km and Vm were about 7 [mu]M and 20 [mu]mol g-1 fresh weight h-1, respectively. Pi uptake in Murashige-Skoog medium under different Pi concentrations and different initial cell densities followed basically the same kinetics. When supplied with abundant Pi, cells absorbed Pi at a constant rate (Vm) for the first hours and accumulated it in the vacuole. As the endogenous pool expanded, the rate of Pi uptake gradually decreased to nil. Maximum Pi accumulation was 100 to 120 [mu]mol g-1 fresh weight if cell swelling during Pi uptake (about 2-fold in cell volume) was not considered. Results indicated that (a) the rate of Pi uptake by Catharanthus cells was independent of initial cell density and was constant over a wide range of Pi concentrations (2 mM to about 10 [mu]M) unless the cells were preloaded with excess Pi, and (b) there was no apparent feedback control over the Pi uptake process in the plasma membrane to avoid Pi toxicity. The importance of the tonoplast Pi transport system in cytoplasmic Pi homeostasis is discussed.