Implication of a positive virtual crossmatch with negative flow crossmatch: A mind-boggler.

There are occasions when tests performed before considering a patient for transplant are ambiguous and require further workup. One such condition is the presence of a positive virtual crossmatch (VXm) (anti-human leukocyte antigen [HLA-A]*26: 01 antibody in this case) with a negative complement-dependent cytotoxicity, Luminex, and flow crossmatch. To ascertain the nature of the antibody, the beads used in single-antigen bead assay (SAB) were treated by acid to denature the antigens and retested with the control and test sample. The mean fluorescence intensities (MFIs) from the patient sera with acid-treated beads increased considerably as compared to the regularly untreated SAB indicating additional antigen epitopes become available by the denaturation process. The MFIs of the antibodies from that of the control sera were reduced to half on testing with the acid-treated SAB assay, indicating that HLA antigen HLA-A*26 was susceptible to acid treatment. Therefore, results of VXm should be interpreted with caution.

Pneumonia Prevention Strategies for Children With Neurologic Impairment.

BACKGROUND: Children with neurologic impairment (NI) face high risk of recurrent severe pneumonia, with prevention strategies of unknown effectiveness. We evaluated the comparative effectiveness of secondary prevention strategies for severe pneumonia in children with NI. METHODS: We included children enrolled in California Children's Services between July 1, 2009, and June 30, 2014, with NI and 1 pneumonia hospitalization. We examined associations between subsequent pneumonia hospitalization and expert-recommended prevention strategies: dental care, oral secretion management, gastric acid suppression, gastrostomy tube placement, chest physiotherapy, outpatient antibiotics before index hospitalization, and clinic visit before or after index hospitalization. We used a 1:2 propensity score matched model to adjust for covariates, including sociodemographics, medical complexity, and severity of index hospitalization. RESULTS: Among 3632 children with NI and index pneumonia hospitalization, 1362 (37.5%) had subsequent pneumonia hospitalization. Only dental care was associated with decreased risk of subsequent pneumonia hospitalization (adjusted odds ratio [aOR]: 0.64; 95% confidence interval [CI]: 0.49-0.85). Exposures associated with increased risk included gastrostomy tube placement (aOR: 2.15; 95% CI: 1.63-2.85), chest physiotherapy (aOR: 2.03; 95% CI: 1.29-3.20), outpatient antibiotics before hospitalization (aOR: 1.42; 95% CI: 1.06-1.92), clinic visit before (aOR: 1.30; 95% CI: 1.11-1.52), and after index hospitalization (aOR: 1.72; 95% CI: 1.35-2.20). CONCLUSIONS: Dental care was associated with decreased recurrence of severe pneumonia. Several strategies, including gastrostomy tube placement, were associated with increased recurrence, possibly due to unresolved confounding by indication. Our results support a clinical trial of dental care to prevent severe pneumonia in children with NI.

Role of de novo DQ donor-specific antibody in antibody-mediated rejection in renal transplant recipient: A case study.

The human leukocyte antigen (HLA) matching plays an important role in determining the clinical outcome of renal transplantation. The development of donor specific antibodies (DSA) against HLA is associated with antibody mediated allograft tissue injury, poor outcome and rejection. The DQ-DSA develops in a denovo pattern and its unfavorable impact on renal transplantation has not yet been widely reported. We investigated the clinical significance of DQ-DSA in a patient diagnosed with hypertension, CKD stage V on maintenance hemodialysis (MHD) for second renal transplant. The histocompatibility workup before the first transplant included low resolution HLA-A, B, DR typing of both patient and donor. HLA type of the patient was HLA-A*29, 68, HLAB*44, 44, DRB1*07, 11. HLA type of the donor was HLA-A*03, 68, HLA-B*39, 44, DRB1*07, 10 with a 3/6 match. The HLA antibody screen and complement dependent cytotoxicity crossmatch (CDC) were found to be negative. No therapeutic plasma exchanges (TPE) were done during stay and post-transplant the patient was on triple immunosuppressant therapy. After four years the patient was diagnosed with recurrent membranoproliferative glomerulonephritis and second renal transplant was planned, therefore, histocompatibility workup was initiated. HLA antibody screen was found to be positive for HLA class II. Initially only HLA-A, B, DR typing was performed and that too only low resolution, further, high resolution HLA typing was done for HLA-DR and DQ to rule out if these antibodies are de-novo DQ/DR DSA. We analyzed that the patient had developed de-novo DSA against HLA-DRB1* 10:01 (DR10), MFI-2374 and DQB1*06:01 (DQ6), MFI-15315. This study suggests the role of DQ antibodies in determining the graft survival and to highlight the need of HLA DQ typing as a routine of the diagnostic work-up in a solid organ transplant.

A case report of successful renal transplantation in an ABO incompatible patient with a preformed donor-specific antibody and negative CDC human leukocyte antigens crossmatch.

ABO incompatibility and preformed antibodies against the human leukocyte antigen (HLA) are two impermissible barriers to a successful renal transplantation, especially in highly sensitized patient population. With the availability of effective desensitization regimens, good patient and graft outcomes have been reported. As transfusion medicine specialists we report our experience, where patient presented with dual histocompatibility barriers i.e. ABO incompatibility along with preformed donor-specific antibodies (DSA) and negative complement dependent lymphocytotoxicity (CDC) HLA crossmatch. The desensitization strategy followed for our patient included rituximab (375 mg/m2), bortezomib (1.3 mg/m2) and eleven pre-transplant therapeutic plasma exchange (TPE) followed by intravenous immunoglobulin (100 mg/kg per TPE session). Anti-B titer of 1:1 and negative Luminex crossmatch (LumXm) class II DSA (less than 1000 mean fluorescence intensity; MFI), was achieved prior to renal transplantation. Fifteen months post-transplant, patient is doing well with serum creatinine level of 0.8 mg/dL with repeat LumXm class II DSA negative (891 MFI). The desensitization regimen followed proved to be effective in our case.

Role of Anti-MICA Antibodies in Graft Survival of Renal Transplant Recipients of India.

INTRODUCTION: The MIC (MHC class I chain-related) genes are a group of nonclassical MHC genes, located in the MHC class 1 region of chromosome 6. The aim of the present study was to find the prevalence of MHC class 1 chain-related (MICA) alloantibodies in patients undergoing live-related donor renal transplantation and its role in short-term graft survival. The role of blood transfusion in the formation of these antibodies was also studied. MATERIALS AND METHODS: Pretransplant samples of patients undergoing renal allograft transplantation were tested for anti-MICA antibodies. Association of various demographics, HLA-A + B + DRB1 mismatches, anti-HLA antibody screen, and anti-MICA antibodies was assessed using Pearson's chi-square test. RESULTS: Out of 646 serum samples, 94 (14.6%) were positive and 552 (85.4%) were negative for anti-MICA antibodies. Patients with anti-MICA antibody had a graft survival 89.3% as compared to 94.7% in patients without anti-MICA antibody (P < 0.05). The hazard ratio for all patients was 3.0701 (P < 0.05). Out of the 340 patients with no HLA antibodies, the presence of anti-MICA antibodies without any HLA antibodies (n = 43) was associated with poor outcome in the patients (hazard ratio of 2.768, P < 0.05). The presence of MICA antibodies with HLA antibodies did not decrease the graft survival (hazards ratio of 1.3750, P > 0.05). CONCLUSION: Preformed MICA antibodies independently increase the risk of kidney rejection and therefore recommend that guidelines should be formed for mandatory testing of these antibodies prior to renal transplant.

A positive complement dependent cytotoxicity immunoglobulin G crossmatch due to auto-antibodies with a negative luminex bead assays in a renal transplant recipient: A Diagnostic dilemma.

Transplant recipients are always at a risk of developing anti-human leukocyte antigen (HLA) antibodies due to prior sensitizing events such as blood transfusions, multiple pregnancies, or transplantation. Unexpected positive outcomes can be seen in complement dependent cytotoxicity (CDC) based assays due to underlying autoimmune disorders or pharmacological treatment (rituximab/intravenous immunoglobulin/anti-thymocyte globulin administration), therefore, limiting its value. CDC based assay results strongly depend on the vitality of the donor lymphocytes, highlighting another major limitation of this assay. Thus, as an alternative approach, solid phase based crossmatch assays were introduced which function independently of the cell quality and have higher sensitivity and specificity in detecting anti-HLA antibodies. We describe a case where the patient awaiting renal transplantation from living related donor was evaluated by pretransplant histocompatibility testing for the detection of anti-HLA antibodies. The histocompatibility testing revealed positive CDC anti-human globulin and flow crossmatch along with negative Luminex based assays (HLA antibody screen, luminex crossmatch, and luminex single bead assay). Detailed histocompatibility workup revealed immunoglobulin G autoantibodies which were complement activating and lympocytoxic in nature.

The Superior Labial Frenulum in Newborns: What Is Normal?

Introduction and Objectives: There has been an emergence of procedures to release the superior labial frenula in infants, yet little is known about the normal appearance or incidence of severe attachment, or "lip-tie." The objective of this article was to develop a classification system for superior labial frenula and to estimate the incidence of different degrees of attachment. Methods: A prospective cross-sectional study. Newborns were examined and had photographs taken of their upper frenula. Relevant medical professionals rated the appearance of the labial frenula using a previously described Kotlow classification system. The raters assessed each photograph twice and were blinded to their previous rating and to other raters' scores. Results: All newborns have a labial frenula, with most attached at the gingival margins (83%). Raters had poor intra- and interrater reliability (64% to 74% and 8%, respectively), using the Kotlow classification system, which improved when the classification system was simplified. Conclusions: The Kotlow classification of lip-tie fails to be reproducible by relevant experts. The majority of infants had a significant level of attachment of the labial frenulum. As more procedures are done to release the upper lip frenulum, it is important to understand what degree of attachment is normal, or more common.

Caries management by risk assessment in a cleft and craniofacial center.

Patients with craniofacial anomalies have an increased incidence of dental caries. The prevention program "Caries Management By Risk Assessment" (CAMBRA) has been previously validated but has not yet been introduced at a widespread level in a medical setting, particularly for this high-risk population.In this cross-sectional study, we aimed to evaluate the feasibility of implementing CAMBRA during the medical visit at an institutional tertiary care center, which treats children with craniofacial anomalies. The study included 161 participants aged 1 to 18 years. Patients and parents received a personalized educational session, toothbrushing tutorial, and fluoride varnish application. We assessed the prevalence of dental caries, caries risk factors, and knowledge of oral hygiene in this patient population.The overall caries prevalence in this group was higher than average (57% compared with 42%, according to the Centers for Disease Control and Prevention). The most prevalent risk factors were developmental delay, deep pits/fissures, low socioeconomic status, orthodontic appliances, and carbohydrate snacks. The greatest predictors of dental caries were having 1 or more risk factors and having low socioeconomic status. In summary, children with craniofacial anomalies were at high risk for dental caries, with high rates of risk factors and low rates of preventive factors.Our findings revealed that basic oral hygiene standards are not being met in this high-risk population, highlighting the need for implementation of protocols such as CAMBRA. The results of this study can aid healthcare workers in craniofacial centers and children's hospitals to improve the understanding of oral hygiene and dental care of their patients.

Cycloartanes from the gum resin of Gardenia gummifera L.f.

The gum resin exuding from the leaf buds of Gardenia gummifera was investigated. Eight new cycloartane triterpenes, 1-6, 8, and 10, together with two known triterpenes, 25-hydroxycycloart-23-en-3-one (7) and cycloartenone (9), were isolated and identified by extensive NMR spectroscopy. For cycloartenone (9), full NMR assignments are given as these data were not available in the literature. Eight compounds possess a C(3)=O group, two are 3,4-secocycloartanes bearing a free C(3)OOH group; in one of the cycloartanes, gummiferartane-9 (10), ring A occurs as a seven-membered lactone.

Antimicrobial Properties of Achyranthes aspera.

Chloroform and methanol root and shoot extracts of A. aspera showed good amount of antibacterial activity against Klebsiella sp. While pet. Ether (60-80°) root extract showed the activity against B. Substilis only antifungal activity of roots was found in extracts with pet ether, chloroform and methanol against fusarium sp. only. Methanol and Aqueous shoot extracts were weakly active against Pencillium. Phytophothora and Sclerouum sp. Results suggest that extract has significant antibacterial and antifungal activities against tested microorganisms. The present study justified the claimed uses of A. aspera in the traditional system of medicine to treat various infectious diseases.