IL-12Rβ1 deficiency in two of fifty children with severe tuberculosis from Iran, Morocco, and Turkey

Background and Objectives

In the last decade, autosomal recessive IL-12Rβ1 deficiency has been diagnosed in four children with severe tuberculosis from three unrelated families from Morocco, Spain, and Turkey, providing proof-of-principle that tuberculosis in otherwise healthy children may result from single-gene inborn errors of immunity. We aimed to estimate the fraction of children developing severe tuberculosis due to IL-12Rβ1 deficiency in areas endemic for tuberculosis and where parental consanguinity is common.

Methods and Principal Findings

We searched for IL12RB1 mutations in a series of 50 children from Iran, Morocco, and Turkey. All children had established severe pulmonary and/or disseminated tuberculosis requiring hospitalization and were otherwise normally resistant to weakly virulent BCG vaccines and environmental mycobacteria. In one child from Iran and another from Morocco, homozygosity for loss-of-function IL12RB1 alleles was documented, resulting in complete IL-12Rβ1 deficiency. Despite the small sample studied, our findings suggest that IL-12Rβ1 deficiency is not a very rare cause of pediatric tuberculosis in these countries, where it should be considered in selected children with severe disease.

Significance

This finding may have important medical implications, as recombinant IFN-γ is an effective treatment for mycobacterial infections in IL-12Rβ1-deficient patients. It also provides additional support for the view that severe tuberculosis in childhood may result from a collection of single-gene inborn errors of immunity.     

Evaluation of clinical outcomes,laboratory and imaging data of patients with solid tumor infected with COVID-19 infection

Background: COVID-19 is associated with higher mortality rates in patients with cancer. In this study, we aimed to evaluate the clinical outcomes, and laboratory and imaging data of patients with solid tumor infected with COVID-19 infection. Methods: This is a cross-sectional retrospective study performed in 2020-2022 on 85 patients with a previous diagnosis of solid tumors infected with COVID-19. We included all patients with tumors of solid organs that were diagnosed with COVID-19 infection and required hospitalization those patients previously hospitalized for treatments and were infected with COVID-19 during hospitalization. Demographic data of patients were collected using a checklist. We collected data regarding clinical outcome (discharge, hospitalization or death), duration of hospitalization, requiring ICU admission, duration of hospitalization divided by received drugs and type of tumor and mean survival time. Furthermore, we collected laboratory data from all patients. The radiologic characteristics of patients were also extracted from their data. Results: Breast cancer was the most common solid tumor (34.9%), followed by lung cancer (19.3%). The mortality rate was 24.1% (20 patients). The highest mortality rate in this study was for metastatic intestinal cancer to the lung (100%, one patient), followed by metastatic prostatic cancer to lung (50%, three patients). The highest hospitalization duration was for patients with glioblastoma multiform (GBM) (30 days). The mean survival time among patients with mortality was 19.15±1.80 days. The mean CT severity score of all patients was 27.53±22.90. Patient’s most common radiologic sign was air space consolidation (89.1%). The highest CT severity score was found in patients with stomach cancer (46.67±5.77). Conclusion: The mortality rate in this study was 24.1%. Based on the results of our study and previous research, special care should be provided to patients with solid tumors during the COVID-19 pandemic and in infected cases.     

Contribution of basic residues of the autolysis loop to the substrate and inhibitor specificity of factor IXa

The autolysis loop (residues 143-154 in chymotrypsinogen numbering) plays a pivotal role in determining the macromolecular substrate and inhibitor specificity of coagulation proteases. This loop in factor IXa (FIXa) has 3 basic residues (Arg143, Lys147, and Arg150) whose contribution to the protease specificity of factor IXa has not been studied. Here, we substituted these residues individually with Ala in Gla-domainless forms of recombinant factor IX expressed in mammalian cells. All mutants exhibited normal amidolytic activities toward a FIXa-specific chromogenic substrate. However, Arg143 and Lys147 mutants showed a approximately 3- to 6-fold impairment in FX activation, whereas the Arg150 mutant activated factor X normally both in the absence and presence of factor VIIIa. By contrast, Arg143 and Lys147 mutants reacted normally with antithrombin (AT) in both the absence and presence of the cofactor, heparin. However, the reactivity of the Arg150 mutant with AT was impaired 6.6-fold in the absence of heparin and 33- to 70-fold in the presence of pentasaccharide and full-length heparins. These results suggest that Arg143 and Lys147 of the autolysis loop are recognition sites for FX independent of factor VIIIa, and Arg150 is a specific recognition site for AT that can effectively interact with AT only if the serpin is in the heparin-activated conformation.     

A multibody modelling approach to determine load sharing between passive elements of the lumbar spine

The human spinal segment is an inherently complex structure, a combination of flexible and semi-rigid articulating elements stabilised by seven principal ligaments. An understanding of how mechanical loading is shared among these passive elements of the segment is required to estimate tissue failure stresses. A 3D rigid body model of the complete lumbar spine has been developed to facilitate the prediction of load sharing across the passive elements. In contrast to previous multibody models, this model includes a non-linear, six degrees of freedom intervertebral disc, facet bony articulations and all spinal ligaments. Predictions of segmental kinematics and facet joint forces, in response to pure moment loading (flexion–extension), were compared to published in vitro data. On inclusion of detailed representation of the disc and facets, the multibody model fully captures the non-linear flexibility response of the spinal segment, i.e. coupled motions and a mobile instantaneous centre of rotation. Predicted facet joint forces corresponded well with reported values. For the loading case considered, the model predicted that the ligaments are the main stabilising elements within the physiological motion range; however, the disc resists a greater proportion of the applied load as the spine is fully flexed. In extension, the facets and capsular ligaments provide the principal resistance. Overall patterns of load distribution to the spinal ligaments are in agreement with previous predictions; however, the current model highlights the important role of the intraspinous ligament in flexion and the potentially high risk of failure. Several important refinements to the multibody modelling of the passive elements of the spine have been described, and such an enhanced passive model can be easily integrated into a full musculoskeletal model for the prediction of spinal loading for a variety of daily activities.     

Association of Mycobacterium infections in patients with Mendelian susceptibility to mycobacterial disease with venous thromboembolism

An association between a hypercoagulable state and Mendelian susceptibility to mycobacterial disease (MSMD) has been established in a few studies; resultant thrombosis is considered rare. In a case‐control study, the prevalence of factor V Leiden, prothrombin G20210A and methylenetetrahydrofolate reductase (MTHFR) C677T, A1298C mutations were investigated in mycobacterium‐infected patients. The study comprised 30 patients with mycobacterial infections (invasive, disseminated and/or recurrent infections with Bacille Calmette–Guerin or non‐tuberculosis mycobacteria and Mycobacterium Tuberculosis with positive results for acid‐fast bacilli and tuberculin skin tests) and 30 normal healthy controls. Forty female (66.7%) and 20 male subjects (33.3%) aged from 3 to 70 years were recruited into this study. Genotyping of targeted genes was performed by RT‐PCR and cytokine TNF‐α concentrations were quantified using a commercially available ELISA kit. Significant associations between mycobacterial infection and TNF‐α production after stimulating peripheral blood mononuclear cells with LPS alone and with IFN‐γ plus LPS were identified. Moreover, genotyping analysis in the studied population revealed a significant association between MTHFR c.677C>T (OR, 3.28; 95% CI, 1.35–7.92; P < 0.05), MTHFR c.1298A>C (OR, 2.33; 95% CI, 1.10–4.93; P < 0.05) and mycobacterial infection in affected patients, indicating susceptibility to venous thromboembolism according to previous studies. Additionally, mycobacterium‐infected patients had a significantly greater prevalence of MTHFR C677T and A1298C mutations than controls.     

Motion repulsion arises from stimulus statistics when analyzed with a clustering algorithm

Motion repulsion is the perceived enlargement of the angle between the directions of motion of two transparently moving patterns. An explanation of this illusion has long been sought for in the neural circuitry of the brain. We show that motion repulsion already arises from the statistical properties of the motion transparency problem when analyzed with a clustering algorithm.     

Identification of noncanonical melanoma-associated T cell epitopes for cancer immunotherapy

The identification of tumor-associated T cell epitopes has contributed significantly to the understanding of the interrelationship of tumor and immune system and is instrumental in the development of therapeutic vaccines for the treatment of cancer. Most of the known epitopes have been identified with prediction algorithms that compute the potential capacity of a peptide to bind to HLA class I molecules. However, naturally expressed T cell epitopes need not necessarily be strong HLA binders. To overcome this limitation of the available prediction algorithms we established a strategy for the identification of T cell epitopes that include suboptimal HLA binders. To this end, an artificial neural network was developed that predicts HLA-binding peptides in protein sequences by taking the entire sequence context into consideration rather than computing the sum of the contribution of the individual amino acids. Using this algorithm, we predicted seven HLA A*0201-restricted potential T cell epitopes from known melanoma-associated Ags that do not conform to the canonical anchor motif for this HLA molecule. All seven epitopes were validated as T cell epitopes and three as naturally processed by melanoma tumor cells. T cells for four of the new epitopes were found at elevated frequencies in the peripheral blood of melanoma patients. Modification of the peptides to the canonical sequence motifs led to improved HLA binding and to improved capacity to stimulate T cells.     

Development and validation of a rapid HPLC method for simultaneous determination of tramadol, and its two main metabolites in human plasma

Tramadol, an analgesic agent, and its two main metabolites O-desmethyltramadol (M1) and N-desmethyltramadol (M2) were determined simultaneously in human plasma by a rapid and specific HPLC method. The sample preparation was a simple extraction with ethyl acetate. Chromatographic separation was achieved with a Chromolith Performance RP-18e 50 mm x 4.6 mm column, using a mixture of methanol:water (13:87, v/v) adjusted to pH 2.5 by phosphoric acid, in an isocratic mode at flow rate of 2 ml/min. Fluorescence detection (lambda(ex)=200 nm/lambda(em)=301 nm) was used. The calibration curves were linear (r(2)>0.997) in the concentration range of 2.5-500 ng/ml, 1.25-500 ng/ml and 5-500 ng/ml for tramadol, M1 and M2, respectively. The lower limit of quantification was 2.5 ng/ml for tramadol, 1.25 ng/ml for M1 and 5 ng/ml for M2. The within- and between-day precisions in the measurement of QC samples at four tested concentrations were in the range of 2.5-9.7%, 2.5-9.9% and 5.9-11.3% for tramadol, M1 and M2, respectively. The developed procedure was applied to assess the pharmacokinetics of tramadol and its two main metabolites following administration of 100mg single oral dose of tramadol to healthy volunteers.