Development of ISB 1442, a CD38 and CD47 bispecific biparatopic antibody innate cell modulator for the treatment of multiple myeloma.

Antibody engineering can tailor the design and activities of therapeutic antibodies for better efficiency or other advantageous clinical properties. Here we report the development of ISB 1442, a fully human bispecific antibody designed to re-establish synthetic immunity in CD38+ hematological malignancies. ISB 1442 consists of two anti-CD38 arms targeting two distinct epitopes that preferentially drive binding to tumor cells and enable avidity-induced blocking of proximal CD47 receptors on the same cell while preventing on-target off-tumor binding on healthy cells. The Fc portion of ISB 1442 is engineered to enhance complement dependent cytotoxicity, antibody dependent cell cytotoxicity and antibody dependent cell phagocytosis. ISB 1442 thus represents a CD47-BsAb combining biparatopic targeting of a tumor associated antigen with engineered enhancement of antibody effector function to overcome potential resistance mechanisms that hamper treatment of myeloma with monospecific anti-CD38 antibodies. ISB 1442 is currently in a Phase I clinical trial in relapsed refractory multiple myeloma.

VPS13D-based disease: Expansion of the clinical phenotype in two brothers and mutation diversity in the Turkish population.

VPS13D is a recently described gene. Worldwide, only 15 families with 23 affected individuals have been reported with a VPS13D-based disease. Mutated VPS13D causes a complex phenotype with a hyperkinetic movement disorder and ataxia, especially in childhood onset disease. The clinical phenotype of the rare adult-onset cases consists of cerebellar ataxia and/or spastic paraplegia. Here, we report the extensive clinical, laboratory and genetic findings of two offspring from consanguineous parents, with ages of disease onset at 57 and 49 with VPS13D-based ataxia. Although conventional magnetic resonance imaging showed mild cerebellar and cerebral atrophy, diffusion tensor imaging, applied for the first time for VPS13D patients, revealed prominent atrophy in U fibers and cerebellopontine tracts. Whole exome sequencing analysis revealed a biallelic Ala4210Val mutation in the VPS13D, reported only once in the literature. Complementary screening of our in-house database consisting of 295 ataxia and hereditary spastic paraplegia patients revealed two further ataxia patients with novel VPS13D variants. Screening the control cohort for VPS13D variants revealed one asymptomatic individual carrying a novel VPS13D variant. In this study, the phenotypic spectrum of VPS13D-based disease is expanded with the description of pre-senile onset predominant ataxia. Further, with the additional novel mutations described, the report is expected to contribute to the understanding of the yet elusive phenotype-genotype correlations in the rare VPS13D-based movement disorder.

Generic high-capacity protein capture and release by pH control.

Techniques for immobilization and release of proteins are of general interest but challenging to develop. Here we show a new method for high-capacity (several µg cm-2) immobilization of proteins in polyelectrolyte brushes by multivalent hydrogen bonds. Upon increasing pH, the proteins are fully released with preserved structure and activity.

Silibinin and ellagic acid increase the expression of insulin receptor substrate 1 protein in ultraviolet irradiated rat skin.

Daily exposure to ultraviolet (UV) light induces inflammation and tumorigenesis in the skin. Silibinin and ellagic acid are natural products that exhibit anti-inflammatory and anti-tumorigenic properties. Insulin receptor substrate protein 1 (IRS1) is important for skin homeostasis and physiology, but its activity following UV radiation remains unclear. We investigated the effects of ellagic acid and silibinin on IRS1 expression in ultraviolet A (UVA) and ultraviolet B (UVB) irradiated rat skin. Forty-two female Wistar rats were divided randomly into six groups of seven animals. The dorsal skin of rats was exposed to UVA + UVB, then treated with ellagic acid and silibinin by gavage. IRS1 expression in skin tissues was determined by western blot analysis. IRS1 expression increased significantly following treatment with ellagic acid and silibinin in UVA + UVB irradiated skin compared to the UVA + UVB only group. After UVA + UVB treatment, ellagic acid effected greater induction of IRS1 expression than silibinin. Our findings suggest that the photoprotective roles of ellagic acid and silibinin may be due to induction of IRS1 expression in UVA + UVB treated rat skin.

Successful Treatment of Renal Infiltration Complicated by Atypical Hemolytic Uremic Syndrome in a Leukemic Child.

Renal infiltration in children with acute leukemia has been reported previously; however, it has rarely been described in association with atypical hemolytic uremic syndrome (aHUS). We present a case of 9-year-old boy who developed life-threatening aHUS in the 1st week of Burkitt leukemia/lymphoma diagnosis with renal infiltration. Complete resolution of aHUS was achieved after therapeutic plasma exchange. This is an uncommon complication of Burkitt leukemia/lymphoma in a pediatric case.

Aza-capped cyclodextrins for intra-cavity metal complexation.

Aza-capped, methylated cyclodextrins (CDs) were obtained in high yields by reacting the soft nitrogen nucleophile 2-nitrobenzenesulfonamide with either A,B-dimesylated CDs in basic media or their diol analogues under Mitsunobu reaction conditions followed by deprotection with thiophenol. A methyl pyridine substituent was grafted on the N atom of these secondary amines. When built on an α-CD scaffold, the resulting tertiary amine no longer undergoes nitrogen inversion at room temperature and behaves as a confining ligand, opening the way to intra-cavity metal complexation and promoting the formation of supramolecular helices.

Impact of glucocorticoid receptor gene (NR3C1) polymorphisms in Turkish patients with metabolic syndrome.

BACKGROUND: The metabolic syndrome (MetS) is characterized by a cluster of metabolic factors, including insulin resistance and type-2 diabetes, abdominal obesity, dyslipidemia, hypertension and microalbuminuria. Impaired glucocorticoid receptor (GR) activity also plays an important role in the etiology of MetS. The objective of our study is to evaluate the effects of GR gene polymorphisms (BclI, N363S, TthIII1 and ER22/23EK) in Turkish patients with MetS. MATERIALS AND METHODS: Seventy subjects with MetS and 185 healthy controls were enrolled in the study. PCR-RFLP analysis was used for genotyping. Results for each polymorphism have been verified by allele-specific oligonucleotide analysis. RESULTS: BclI GG genotype was significantly associated with an increased risk of MetS (p = 0.02). Also, only in women, the G allele carriers were significantly associated with higher C-peptide. T allele carriers of TthIII1 polymorphism were significantly associated with higher C-peptide, triglyceride, insulin and C-reactive protein (CRP, p value 0.048, 0.022, 0.005 and 0.022, respectively), and lower fasting blood glucose (FBG, p = 0.02). The combined carriers of BclI polymorphism G allele and TthIII1 polymorphism T allele were significantly associated with higher diastolic blood pressure in all patients, and lower FBG and postprandial blood glucose in only men. All the ER22/23EK polymorphisms coexisted with polymorphic variant of TthIII1 (p = 0.0058). CONCLUSION: The presence of homozygote polymorphic variant of BclI might be good predictive markers for the disease susceptibility. The BclI and the TthIII1 polymorphism are associated with sex-specific clinical parameters. Our findings also suggest that the combination of BclI and TthIII1 polymorphisms may play a protective role in blood glucose.

Prevalence of sleep disturbance in chronic pain.

OBJECTIVE: Sleep is a vital function for human beings, which can be affected by several factors. Chronic pain is one of these factors where it is the most frequent cause for seeking medical care in combination with insomnia. The aim of this study is to examine the prevalence and relationship between sleep disturbance and chronic pain. PATIENTS AND METHODS: After approval, a total of 85 Family Medicine Units from over 170 in Tokat were randomly selected using a 50% sampling. A sample of 2635 subjects, over the age of 19 years, who were registered with the selected Family Medicine Units, were assessed due to gender, age group, and the urban/rural population size of Tokat using the stratified sampling method. The sample size distribution was calculated as 1515 urban subjects, 1120 rural subjects; 1345 female subjects, 1290 male subjects; 1123 subjects between 20-39 years of age, 1103 subjects between the ages of 40-64, and 409 subjects over 64 years of age. After sampling, subjects were invited to participate in the study via an invitation letter, and agreeing individuals were taken to the Family Medicine Unit for face-to-face meetings. Written, informed consent was obtained, along with demographic data. The presence of chronic pain was recorded. According to the presence of chronic pain, all subjects were separated into two groups as Group Chronic Pain and Group Non-Chronic Pain. The visual analog scale for pain intensity, and Pittsburgh Sleep Quality Index for sleep quality, were performed with all subjects. A multiple linear regression model was used to assess the predictors of sleep quality. Analyses were conducted using the Statistical Package for Social Sciences program (SPSS Inc., Chicago, IL, USA), version 20.0. The statistical significance for all analyses was set at p < 0.05. RESULTS: The mean global Pittsburgh Sleep Quality Index score of Group Chronic Pain (5.30 ± 4.29) was significantly higher than in Group Non-Chronic Pain (3.22 ± 3.30; p < 0.01). The mean Pittsburgh Sleep Quality Index scores of females (5.69 ± 4.40) were significantly higher than males (4.54 ± 3.96) in Group Chronic Pain (p = 0.000045). A total of 40.7% of patients in Group Chronic Pain, and 21.9% in Group Non-Chronic Pain demonstrated poorer sleep quality according to the Pittsburgh Sleep Quality Index scores, with a cut-off level > 5. A moderate positive correlation was found between the global Pittsburgh Sleep Quality Index and Visual Analog Scale scores (r = 0.310, p < 0.01). A multiple linear regression analysis showed that age, gender, income, Visual Analog Scale, and presence of depression were the significant predictors for Pittsburgh Sleep Quality Index score. CONCLUSIONS: The current study revealed that chronic pain and pain intensity are important predictors of sleep quality.

Central nervous system toxicity after acute oral formaldehyde exposure in rabbits: An experimental study.

Formaldehyde (FA) is one of the most widely used chemical compounds in industrial field. It is described as toxic, particularly to the nervous system, the urogenital system, and the respiratory tracts. In this study, we determined the effects of acute oral exposure to FA in rabbit brain tissue. A total of 16 rabbits were selected and divided into 2 groups: formaldehyde group (group F) and control group (group C). FA was administered to group F at a rate of 40 mg/kg/day via a nasogastric tube for 5 days. Saline was similarly administered to the eight controls. All the animals were euthanized after 5 days of exposure, and brain tissue samples were collected in 10% neutral formalin and embedded in paraffin. To investigate the effects of FA on the apoptotic process, we examined active caspase-3, Bax, and Bcl-2 immunohistochemical expression and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate -biotin nick-end labeling (TUNEL) reactivity in the rabbit brains. In addition, glial fibrillary acidic protein (GFAP) was biochemically assessed in brain tissue samples for neurotoxicity. We found that FA treatment caused a significant decrease in Bcl-2 expression and an increase in active caspase-3 and Bax expressions as well as an increase in the number of TUNEL-positive apoptotic cells. The GFAP level was found to be significantly higher in group F. In conclusion, acute oral exposure to FA caused DNA damage, apoptosis, and neuronal injury in the rabbit brains.

Association of serum paraoxonase activity and coronary artery calcification.

AIM: It has been known that there was a relation between the activities of serum paraoxonase (PON) and the severity of the coronary artery disease. However, little is known about association of coronary artery calcification (CAC) and serum PON activities. The aim of this study is to evaluate the relationship between CAC and serum PON activities. PATIENTS AND METHODS: We measure serum PON activities from 122 patients (42 female, mean age = 62±10 years) with angiographically documented CAC (Group I), and 138 patients (54 female, mean age = 60±10 years) without CAC (Group II). Coronary calcification was detected with fluoroscopy before coronary angiography. Serum PON activities were measured by spectrophotometrically method. Patient characteristics and baseline data were recorded from patient's files. RESULTS: The triglyceride levels is lower in group I than group II (p = 0.040). Diastolic blood pressure and frequency of diabetes mellitus was higher in the group I than group II (respectively p = 0.012 and p = 0.022). The other clinic and laboratory parameters were similar in two groups (all p > 0.05). The only statistically significant differences between with CAC and without CAC groups in respect to serum PON activities were present (170.6 ± 59.6 vs. 209.6 ± 69.8 U/ml; respectively, p < 0.001). There was a negative correlation between serum PON activities and presence of CAC (p < 0.001). CONCLUSIONS: These data indicate that the serum PON activities are decreased in patients with CAC. The serum PON activities may play a role in development of the CAC and reduced serum PON activity might represent a biochemical marker of CAC.

Serum ceruloplasmin levels in acute decompensated heart failure.

BACKGROUND: Ceruloplasmin (Cp) is a serum protein that belongs to the family of α2-globulins and it is increased in patients with after acute myocardial infarction complicated with heart failure. Aim of the study was to investigate levels of serum Cp in patients with acute decompensated heart failure. MATERIALS AND METHODS: The present cross-sectional observational study consists of three groups: Fifty patients with decompensated heart failure (Group 1) and same 50 patients after compensation (Group 2); 50 control patients group with comparable age and sex without heart failure (Group 3). Demographic, echocardiographic and biochemical data of patients were collected. Serum Cp level was determined spectrophotometrically. RESULTS: Serum ceruloplasmin was significantly increased in Groups 1 (820.8 ± 78.5 IU/dL) and 2 (873.5 ± 121.0 IU/dL) compared, to Group 3 (640.6 ± 132.4 IU/dL) (p<0.001). In the sub-group analysis, this difference was due to the difference between Groups 3, Group 1 and 2 (both p=0.0001) whereas no significant difference was present between Group I and Group 2 (p>0.063). A positive correlation was found between Cp and female sex, heart rate, systolic and diastolic blood pressure, acetylsalisilic acid and diuretic use, left ventricular systolic and diastolic diameter, mitral regurgitation, and negative correlation was found between Cp and ejection fraction (p<0.05 for all) whereas none of the parameters were independently associated with serum Cp level (p>0.05). CONCLUSIONS: Findings of the present study suggest that serum Cp level is increased in both decompensated and compensated HF compared to controls. Further large scale studies are needed to elucidate the pathophysiological mechanisms of increased Cp in HF.

Effects of isoflurane in an intoxication model: experimental study.

BACKGROUND: Isoflurane is a volatile anaesthetic that has been commonly used since 1980. The major metabolites of isoflurane are fluoride ion and trifluoroacetate, both excreted in the urine. AIM: This study manage to show the histopathological findings of ingested isoflurane on liver, kidney and lugs in an animal model. Twenty-one rabbits were selected and divided into three groups: Group Isoflurane-5 (I-5); Group Isoflurane-10 (I-10); and Group Control (C). Each group consisted of seven rabbits. I-5 and I-10 received 5 ml/kg and 10 ml/kg of liquid isoflurane, respectively, via nasogastric tube, while C received 5 ml/kg saline (0.9% NaCI). All animals in I-5 and I-10 were sacrificed without anesthetic drug administration. Tissue samples from livers, kidneys and lungs were collected, preserving tissue unity and avoiding infliction of any trauma. Samples were fixed in 10% formalin solution, embedded in paraffin blocks and sliced into 5 µm sections. To investigate the effects of isoflurane, sections were examined under light microscope and histopathological changes were scored. RESULTS: Mean injury scores and the appearance of portal lymphocyte infiltration in liver samples showed significant increases in I-5 and I-10 compared to C (p = 0.005, p = 0.001 and p = 0.001, respectively). Mean lung injury scores revealed significant increases after isoflurane treatment in I-5 and I-10 compared to C (p = 0.026 and p = 0.017, respectively). CONCLUSIONS: Ingested isoflurane led to mild liver and lung injuries in rabbits.

Delayed right atrial lateral electromechanical coupling relative to the septal one can be associated with paroxysmal atrial fibrillation.

BACKGROUND: Non-invasive prediction of paroxysmal atrial fibrillation (PAF) is one of the most recent interests of cardiology. AIM: The current study investigates the relationship between the atrial electromechanical coupling time (EMCT) and PAF. PATIENTS AND METHODS: A group of 35 patients with PAF was compared with a group of 37 subjects without PAF. Pulsed wave tissue Doppler evaluations of atrial walls were performed from apical four chambers view under ECG monitoring. The time intervals from the onset of P wave to the onset of late diastolic wave (A') at right atrial wall (P-RA), interatrial septum (P-IAS), and left atrial wall (P-LA, maximum EMCT) were measured. The right atrial EMCT (P-RA minus P-IAS), left atrial EMCT (P-LA minus P-IAS) and interatrial EMCT (P-LA minus P-RA) were computed. A' wave velocities were measured from each atrial wall. RESULTS: RA (16.0±13.1 vs. -8.7±18.6 ms, p < 0.001) and maximum (91.5±32.6 vs. 72.0±23.1 ms, p = 0.001) EMCT were longer, RA A' velocity was higher in the patient group. There were no differences between the groups in LA and interatrial EMCT, and septal and LA A' velocities. Regression analysis revealed that only RA [OR: 1.148 (1.041-1.267), p = 0.006] and maximum [OR: 1.099 (1.009-1.197), p = 0.031] EMCT were independent variables for PAF. In order to predict patients with PAF, we have chosen +7.5 msn for the RA EMCT which yielded 69% sensitivity and 71.4% specificity to predict patients. CONCLUSIONS: Delayed RA lateral EMCT relative to septal one and delayed maximum EMCT detected by tissue Doppler could be a valuable method for identifying patients with PAF.

GMP-adapted overexpression of CXCR4 in human mesenchymal stem cells for cardiac repair.

BACKGROUND: Human mesenchymal stem cells (MSC) have been utilized for cardiac regeneration after myocardial damage. Their clinical effects are marginal and only a minority of administered cells could make their way into the myocardium. The chemokine receptor CXCR4 has been identified as crucial for migration and homing of stem cells. In this study we overexpressed CXCR4 on human MSC to improve cell trafficking and tissue repair. METHODS: Human MSC were isolated from the spongiosa of tibia and femur as well as from pelvic bone marrow. MSC were characterized by differentiation assays and FACS analysis. CXCR4 was overexpressed by mRNA-nucleofection. Intracellular signaling was analyzed to demonstrate functionality of CXCR4. The modified Boyden chamber, wounding assays and time lapse microscopy were utilized to investigate MSC migration. RESULTS: MSC did not express relevant amounts of CXCR4 spontaneously. CXCR4 could be overexpressed in 93% of MSC with a cell viability of 62%. Functionality of the overexpressed CXCR4 was demonstrated by a significant cytosolic Ca(2+) increase and activation of different MAP kinases followed by SDF-1α stimulation. In contrast no improvement of cell migration could be observed. There was a strong basal MSC chemokinesis independent from CXCR4 expression. CONCLUSIONS: CXCR4 could be effectively overexpressed in human MSC by mRNA-nucleofection. Despite functionality of CXCR4 MSC were characterized by a strong basal chemokinesis that could not be further enhanced by CXCR4 overexpression. As isolation, culture and nucleofection of pelvic bone marrow-derived MSC basically fulfill the GMP-requirements our approach seems suited for an in vivo application in patients.

Prospective, randomized, double-blinded comparison of the effects of caudally administered levobupivacaine 0.25% and bupivacaine 0.25% on pain and motor block in children undergoing circumcision surgery.

BACKGROUND: Caudal anesthesia is widely used as intraoperative and postoperative analgesia in children's subumbilical surgeries such as on the urogenital system, lower extremities and lower abdomen to reduce the stress response to surgery and to facilitate the general anesthesia. AIM: The purpose of this study was to compare the effects of caudally administered bupivacaine and levobupivacaine of equal volume and concentration on motor block and postoperative pain in children undergoing circumcision surgery. PATIENTS AND METHODS: The prospective, randomized, double-blind study included 60 patients with ages ranging from 1-10 years and ASA (American Society of Anesthesiologists) physical status of I-II who underwent elective circumcision surgery. The patients were divided into two groups: group B received 0.5 ml/kg of bupivacaine 0.25% caudally and group L received 0.5 ml/kg of levobupivacaine 0.25% caudally. Postoperative pain was assessed by children's and infant's postoperative pain scale and motor block was assessed by the Bromage scale. RESULTS: The mean children's and infant's postoperative pain scale of group B was significantly lower than that of group L (p < 0.001). Three patients in group B and seven patients in group L needed additional analgesia after the incision. There was no significant difference between groups in terms of Bromage scores and in both groups the residual motor block was found to be zero at the 150th minutes. CONCLUSION: According to these findings, bupivacaine has an adequate quality of analgesia than levobupivacaine. We suggest that bupivacaine for caudal block at the concentration of 0.25% (0.5 ml/kg) provides an adequate level of analgesia for outpatient circumcision surgery.

HMGB1 is an independent predictor of death and heart transplantation in heart failure.

BACKGROUND: High-Mobility-Group Box 1 (HMGB1) has been established as an important mediator of myocardial inflammation and associated with progression of heart failure (HF). The aim of this study was to analyze the prognostic value of systemic HMGB1 levels in HF patients with ischemic and non-ischemic cardiomyopathy. METHODS AND RESULTS: We conducted an analysis (median follow-up time 2.5 years) of HMGB1 plasma concentration in 154 patients with systolic HF and correlated the results with disease severity and prognosis. HMGB1 in HF patients with severe symptoms (NYHA III/IV; 5.35 ng/ml; interquartile range (IQR) = 3.48-8.42 ng/ml) was significantly elevated compared with that in patients with mild symptoms (NYHA I/II; 3.37 ng/ml, IQR = 2.31-5.22 ng/ml, p < 0.0001) and with controls (3.25 ng/ml, IQR = 3.04-3.67 ng/ml, p < 0.0001). HMGB1 levels correlated with other markers of heart failure indicating an association of HMGB1 with disease severity in HF. In a univariate cox regression model for the combined endpoint of death and heart transplantation, HMGB1 proved to be a predictor at cut-off values based on HMGB1 terciles of either 3.4 or 6.1 ng/ml (p = 0.001 and p < 0.0001, respectively). In a multivariate cox regression model, which included NT-proBNP, creatinine, age, NYHA class, white blood cell count, anemia, and age, HMGB1 remained an independent predictor of the combined endpoint (hazard ratio (HR) = 2.48, 95% confidence interval (CI) = 1.06-5.83, p = 0.037 and HR = 2.48, 95% CI = 1.31-4.71, p = 0.005, respectively). CONCLUSION: Our findings demonstrate that HMGB1 plasma concentration is elevated in HF and correlates with disease severity and that is an independent predictor of the combined endpoint death and heart transplantation in HF patients.