Applications of flow cytometry in the analysis of blood leukocytes

Flow cytometric analysis of blood leukocytes is currently used for both routine clinical measurements as well as for cutting edge research applications. This technology has enabled rapid and accurate determination of leukocyte antigens and quantitative analysis of leukocyte subsets, tests of leukocyte function, determination of the presence of antineutrophil and antilymphocyte antibodies in plasma and on cells, measurement of CD34+ hematpoietic stem cells in peripheral blood and bone marrow samples, measurement of apoptosis, and detection of virus-infected leukocytes. This review will focus on the use of the flow cytometer for investigations of blood leukocytes in transfusion medicine.     

Role of depressive illness in the outcome of treatment of temporomandibular joint pain-dysfunction syndrome

The aim of this study was to assess the role of a depressive illness in the outcome of the treatment of patients with temporomandibular joint pain-dysfunction syndrome. One group was considered psychiatrically normal and the other had a concurrent depressive illness. The latter group was subdivided equally to produce three treatment groups: one undergoing occlusal splint therapy, one receiving antidepressant medication, and the third having a combination of occlusal splint and antidepressant therapy. The results showed clearly that there was a significant difference in response in the nonpsychiatric and combined-therapy depressed groups in comparison with the two depressed groups treated either with occlusal splint or with antidepressant therapy. The combined therapy led to resolution of the painful problem and the depression, whereas the single therapies were only partly successful in relieving the pain-dysfunction syndrome. The preexisting duration of this painful problem did not influence the response to therapy.     

A 5-year study comparing a posterior composite resin and an amalgam

Eighty class I and class II light-cured posterior composite resin restorations were compared with 43 class I and class II amalgam restorations during a 5-year period after placement. The results of this clinical study showed that both materials were satisfactory during the time period and that the only significant statistical differences are a poorer marginal integrity for the amalgam and a greater wear rate for the composite resin.     

Expression of claudin‐11 by tumor cells in cutaneous squamous cell carcinoma is dependent on the activity of p38δ

The incidence of cutaneous squamous cell carcinoma (cSCC) is rapidly increasing, and the prognosis of patients with metastatic disease is poor. There is an emerging need to identify molecular markers for predicting aggressive behaviour of cSCC. Here, we have examined the role of tight junction (TJ) components in the progression of cSCC. The expression pattern of mRNAs for TJ components was determined with RNA sequencing and oligonucleotide array‐based expression analysis from cSCC cell lines (n=8) and normal human epidermal keratinocytes (NHEK, n=5). The expression of CLDN11 was specifically elevated in primary cSCC cell lines (n=5), but low or absent in metastatic cSCC cell lines (n=3) and NHEKs. Claudin‐11 was detected in cell‐cell contacts of primary cSCC cells in culture by indirect immunofluorescence analysis. Analysis of a large panel of tissue samples from sporadic UV‐induced cSCC (n=65), cSCC in situ (n=56), actinic keratoses (n=31), seborrhoeic keratoses (n=7) and normal skin (n=16) by immunohistochemistry showed specific staining for claudin‐11 in intercellular junctions of keratinizing tumor cells in well and moderately differentiated cSCCs, whereas no staining for claudin‐11 was detected in poorly differentiated tumors. The expression of claudin‐11 in cSCC cells was dependent on the activity of p38δ MAPK and knock‐down of claudin‐11 enhanced cSCC cell invasion. These findings provide evidence for the role of claudin‐11 in regulation of cSCC invasion and suggest loss of claudin‐11 expression in tumor cells as a biomarker for advanced stage of cSCC.     

Evaluation of Evidence for Pathogenicity Demonstrates That BLK,KLF11, and PAX4 Should Not Be Included in Diagnostic Testing for MODY

Maturity-onset diabetes of the young (MODY) is an autosomal dominant form of monogenic diabetes, reported to be caused by variants in 16 genes. Concern has been raised about whether variants in BLK (MODY11), KLF11 (MODY7), and PAX4 (MODY9) cause MODY. We examined variant-level genetic evidence (cosegregation with diabetes and frequency in population) for published putative pathogenic variants in these genes and used burden testing to test gene-level evidence in a MODY cohort (n = 1,227) compared with a control population (UK Biobank [n = 185,898]). For comparison we analyzed well-established causes of MODY, HNF1A, and HNF4A. The published variants in BLK, KLF11, and PAX4 showed poor cosegregation with diabetes (combined logarithm of the odds [LOD] scores ≤1.2), compared with HNF1A and HNF4A (LOD scores >9), and are all too common to cause MODY (minor allele frequency >4.95 × 10⁻⁵). Ultra-rare missense and protein-truncating variants (PTV) were not enriched in a MODY cohort compared with the UK Biobank population (PTV P > 0.05, missense P > 0.1 for all three genes) while HNF1A and HNF4A were enriched (P < 10⁻⁶). Findings of sensitivity analyses with different population cohorts supported our results. Variant and gene-level genetic evidence does not support BLK, KLF11, or PAX4 as a cause of MODY. They should not be included in MODY diagnostic genetic testing.     

Maternal eating disorders influence sex ratio at birth

We explored sex ratio at birth, defined as the proportion of male live births, in women with anorexia nervosa, bulimia nervosa, binge eating disorder, and eating disorders not otherwise specified-purging type (EDNOS-P) relative to a referent group in a large population-based sample of 38,340 pregnant women in Norway. Poisson regressions were adjusted for mother's age, pre-pregnancy BMI, lifetime smoking status, maternal education, income, marital status, gestational age, and parity. Lower proportions of male live births were observed in the anorexia and bulimia groups, while binge eating disorder and EDNOS-P were associated with a higher proportion of male births. These data suggest that maternal eating disorders may influence offspring sex and that the direction of effect may vary by eating disorder subtype. If confirmed, this finding could provide evidence in formulating hypotheses regarding the consequences of eating disorders and determinants of sex ratio at birth.