Ex vivo-expanded human CD19+TIM-1+ regulatory B cells suppress immune responses in vivo and are dependent upon the TIM-1/STAT3 axis.

Regulatory B cells (Breg) are a heterogenous population with immune-modulating functions. The rarity of human IL-10+ Breg makes translational studies difficult. Here we report ex vivo expansion of human B cells with in vivo regulatory function (expBreg). CD154-stimulation of human CD19+ B cells drives >900-fold expansion of IL-10+ B cells that is maintained in culture for 14 days. Whilst expBreg-mediated suppressive function is partially dependent on IL-10 expression, CRISPR-mediated gene deletions demonstrate predominant roles for TIM-1 and CD154. TIM-1 regulates STAT3 signalling and modulates downstream suppressive function. In a clinically relevant humanised mouse model of skin transplantation, expBreg prolongs human allograft survival. Meanwhile, CD19+CD73-CD25+CD71+TIM-1+CD154+ Breg cells are enriched in the peripheral blood of human donors with cutaneous squamous cell carcinoma (SCC). TIM-1+ and pSTAT3+ B cells are also identified in B cell clusters within histological sections of human cutaneous SCC tumours. Our findings thus provide insights on Breg homoeostasis and present possible targets for Breg-related therapies.

Olive oil polyphenol oleuropein inhibits smooth muscle cell proliferation.

OBJECTIVES: The Mediterranean diet, with a high content of olive oil, is associated with a reduced risk of coronary artery disease. The aim of this study was to determine the effect of oleuropein, one of the polyphenols in olive oil, on vascular smooth muscle cell (SMC) proliferation in vitro. DESIGN: This was an experimental study. MATERIALS AND METHODS: Bovine vascular SMCs were cultured in the presence of 100 µM of oleuropein. On days 1, 3 and 5, cell number was counted. Cell cycle analysis was performed by flow cytometry. Cell cycle regulators were assessed by immunoblotting. RESULTS: Cell proliferation in the presence of oleuropein was significantly inhibited by 92%. Cell cycle analysis revealed that oleuropein treatment blocked cells in the G1-S phase compared with the non-treated group. Among G1 phase regulators, retinoblastoma protein, cyclinD, p21 and p27 were not affected by oleuropein, but extracellular signal-regulated kinase 1/2 (ERK1/2) activation was inhibited. Growth of SMC treated with 100 µM of the mitogen-activated protein (MAP) kinase/ERK kinase 1 (MEK1) inhibitor PD98059 was also significantly inhibited by 70%. CONCLUSIONS: Oleuropein inhibited SMC proliferation through a cell cycle block between the G1 and the S phases, which may be regulated by ERK1/2. These results suggest a mechanism by which olive oil consumption may have beneficial effects on cardiovascular mortality by inhibiting SMC proliferation.

Analysis of benefit:risk ratio and mortality reduction for the UK Breast Screening Programme.

A quantitative analysis has been performed to predict the benefit:risk ratio and associated mortality reduction for the UK National Health Service Breast Screening Programme. The analysis is based on the results of an established biological simulation method coupled with dosimetric information and population statistics applicable to the UK breast screening programme. As well as the general breast screening population, the benefit:risk ratios for specific subgroups of women thought to be at higher risk are estimated. The effects of alterations in screening strategy are also investigated. The results indicate favourable benefit:risk ratios and mortality reductions for all women in the programme, with a breast cancer mortality reduction of approximately 9% over the whole UK female population, equivalent to a breast cancer mortality reduction in the region of 25% for the age range 55-69 years.

Psychogenic mania and bereavement.

Early notions of mania invoked a combination of psychogenic and organic factors, but psychogenic mania has not endured as a concept. We present a 37-year-old woman with an acute manic episode precipitated by the prolonged death of her husband from cancer. To our knowledge this is the first published account of 'maniacal grief' in the absence of pre-existing affective disorder, and with clear causal relations. Mania was clearly induced by the bereavement and we argue that the loss represented more than a non-specific life event. This case supports a re-examination of mania as potentially a 'reactive' or psychogenic phenomenon.

Additional factors for the estimation of mean glandular breast dose using the UK mammography dosimetry protocol.

The UK and European protocols for mammographic dosimetry use conversion factors that relate incident air kerma to the mean glandular dose (MGD) within the breast. The conversion factors currently used were obtained by computer simulation of a model breast with a composition of 50% adipose and 50% glandular tissues by weight (50% glandularity). Relative conversion factors have been calculated which allow the extension of the protocols to breasts of varying glandularity and for a wider range of mammographic x-ray spectra. The data have also been extended to breasts of a compressed thickness of 11 cm. To facilitate the calculation of MGD in patient surveys, typical breast glandularities are tabulated for women in the age ranges 40-49 and 50-64 years, and for breasts in the thickness range 2-11 cm. In addition, tables of equivalent thickness of polymethyl methacrylate have been provided to allow the simulation for dosimetric purposes of typical breasts of various thicknesses.

Estimation of mean glandular dose for mammography of augmented breasts.

The standard quantity used to relate breast surface exposure to radiation risk is the mean dose received by the radiation sensitive tissue contained within the female breast, the mean glandular dose (MGD). At present, little is known about the MGD received by women with breast implants as there is no technique available to facilitate its calculation. The present work has involved modification of the conventional method for MGD estimation to make it applicable to women with augmented breasts. The technique was used to calculate MGDs for a cohort of 80 women with breast implants, which were compared with similar data calculated for a total of 1258 non-augmented women. Little difference was found in median MGD at low compressed breast thickness. At high breast thickness, however, the MGDs received by women with augmented breasts were found to be considerably lower than those relating to their non-augmented counterparts.

Dosimetric implications of age related glandular changes in screening mammography.

The UK National Health Service Breast Screening Programme is currently organized to routinely screen women between the ages of 50 and 64, with screening for older women available on request. The lower end of this age range closely matches the median age for the menopause (51 years), during which significant changes in the composition of the breast are known to occur. In order to quantify the dosimetric effect of these changes, radiographic factors and compressed breast thickness data for a cohort of 1258 women aged between 35 and 79 undergoing breast screening mammography have been used to derive estimates of breast glandularity and mean glandular dose (MGD), and examine their variation with age. The variation of mean radiographic exposure factors with age is also investigated. The presence of a significant number of age trial women within the cohort allowed an extended age range to be studied. Estimates of MGD including corrections for breast glandularity based on compressed breast thickness only, compressed breast thickness and age and for each individual woman are compared with the MGD based on the conventional assumption of a 50:50 adipose/glandular composition. It has been found that the use of the conventional 50:50 assumption leads to overestimates of MGD of up to 13% over the age range considered. By using compressed breast thickness to estimate breast glandularity, this error range can be reduced to 8%, whilst age and compressed breast thickness based glandularity estimates result in an error range of 1%.

Negative feedback potency of estradiol is increased in orchidectomized sheep during chronic nutrient restriction.

The effect of nutrient restriction on serum concentrations of the gonadotropins, on the pattern of LH secretion, and on sensitivity to estradiol (E2) was assessed in orchidectomized sheep (wethers). Thirty-six wethers (initial weight = 42.3 +/- 0.6 kg) were fed to gain, maintain, or lose body weight (feeding groups G, M, and L, respectively; n = 12 wethers/group). After 7 wk of controlled feeding, G, M, and L wethers weighed 57.0 +/- 1.7, 42.5 +/- 0.6, and 36.6 +/- 0.8 kg, respectively. At the end of controlled feeding, serum concentrations of LH and FSH in M and L wethers were significantly higher than levels in G wethers. During Days 51-54 of controlled feeding, six animals from each feeding group received E2 or vehicle as a continuous infusion. The pattern of LH secretion was assessed 48 h after initiation of infusion. Although E2 infusion did not affect (p > 0.05) LH pulse frequency in G and M wethers, pulse frequency was reduced (p < 0.05) in L wethers receiving E2. These data indicate that serum concentrations of gonadotropins are increased in wethers fed to maintain or lose body weight. In addition, the negative feedback potency of E2 is enhanced during prolonged weight loss.

Moderate and severe nutrient restriction has divergent effects on gonadotroph function in orchidectomized sheep.

The effect of plane of nutrition and estradiol (E2) on pituitary concentrations of GnRH receptor and GnRH receptor mRNA was assessed in orchidectomized sheep (wethers). As detailed in the companion paper, 36 wethers were fed to gain, maintain, or lose body weight. Six animals from each feeding group received E2 (0.31 microg E2/50 kg per h) or vehicle during Days 51-54 of controlled feeding. Anterior pituitary tissue was collected at the end of infusion. Both moderate and severe nutrient restriction increased (p < 0.05) tissue concentrations of FSH and FSHbeta mRNA. Conversely, concentrations of GnRH receptor and receptor mRNA were not affected (p > 0.05) by plane of nutrition. Estradiol increased (p < 0.05) GnRH receptor and receptor mRNA in wethers fed to gain or maintain weight. However, this E2-induced response was not evident in wethers subject to severe nutrient restriction. These data demonstrate that severe, but not moderate, nutrient restriction suppresses E2-induced augmentation of tissue concentrations of GnRH receptor and GnRH receptor mRNA. Collectively, the data presented here and in the companion paper suggest that severe nutrient restriction leads to physiologic changes that render the hypothalamus increasingly sensitive to estrogenic stimulation, while the pituitary is made less responsive to steroidal inputs.

Duplicated plastid and triplicated cytosolic isozymes of triosephosphate isomerase in maize (Zea mays L.).

We studied electrophoretic variation and inheritance of triosephosphate isomerase (TPI) isozymes in maize (Zea mays L.). In contrast to most diploid plants, in maize, TPI exists as multiple isozymes in both the plastid and cytosolic subcellular compartments. Phenotypes result from the overlay of two independent sets of isozymes and allozymes, representing the plastid (encoded by the nuclear genes Tpi1 and Tpi2) and cytosolic (encoded by Tpi3, Tpi4, and Tpi5) systems. All possible intragenic and intergenic dimeric enzymes are formed between polypeptides within each subcellular compartment. No heterodimers are formed between plastid and cytosolic polypeptides. Extensive surveys of accessions of land races and inbred lines revealed 22 allelic variants for the five loci. Most alleles have been formally validated by segregation analysis. We describe two null alleles at Tpi4, distinguished by their relative abilities to form intergenic heterodimers with polypeptides specified by Tpi3 and Tpi5. Linkage analyses and crosses with B-A translocation stocks were effective in determining the chromosome locations of all five loci. Duplicated genes for both the plastid and cytosolic isozymes were localized to genomic regions that possess numerous other redundant sequences. We placed Tpi1 on the long arm of chromosome 7, approximately 23 centimorgans (cM) distal to g11; we localized its duplicate--Tpi2--17 cM distal to v4 on the long arm of chromosome 2. The triplicate loci encoding cytosolic TPIs reside on chromosomes 3 and 8. Tpi4 is approximately equidistant (11 cM) from d1 and Lg3, near the centromere of chromosome 3. Tpi3 and Tpi5 are located on distal ends of the most poorly marked maize chromosome; Tpi3 is 29 cM distal to Idh 1 on 8L, and Tpi5 is on 8S or near the centromere on 8L. In contrast to most duplicated maize sequences, which often occur in parallel linkages on different chromosomes, Tpi3 and Tpi5 provide an example of intrachromosomal gene duplication. Several of the Tpi loci are located in sparsely mapped regions of the genome, and Tpi1 is the first isozyme marker for chromosome 7.

Immunoperoxidase localization of Sporothrix schenckii and Cryptococcus neoformans. Staining of tissue sections fixed in 4% formaldehyde solution and embedded in paraffin.

An indirect immunoperoxidase staining method has been successfully applied to 4% formaldehyde solution tissue sections fixed in and embedded in paraffin for the localization of Sporothrix schenckil and Cryptococcus neoformans without prior trypsinization of tissue sections. A comparison of this method with an analogous immunofluorescence staining technique has been made.

Immunoperoxidase localization of treponema pallidum: its use in formaldehyde-fixed and paraffin-embedded tissue sections.

The spirochete, Treponema pallidum, the causative agent of syphilis, has been successfully localized in formaldehydefixed and paraffin-embedded tissue sections using rabbit anti-T pallidum antiserum with two immunoperoxidase techniques. These techniques, the indirect peroxidase-labeled antibody method and the peroxidase-antiperoxidase (PAP) method are compared for sensitivity and degree of nonspecific staining. Both offer substantial advantages over conventional silver-impregnation techniques, but the indirect peroxidase-labeled antibody method seems better, based on the intensity of staining and the simplicity of procedure.

Recurring digital fibrous tumors of childhood: a review.

Two new cases that conform to the clinical and histopathological features of recurring digital fibrous tumor of childhood, described by Reye in 1965, are reported. This tumor, which is considered a distinct entity among the juvenile fibromatoses, characteristically presents in infancy and early childhood, involves only the digits, recurs frequently following surgical excision without metastatic spread, and demonstrates the distinct histopathological finding of intracytoplasmic inclusion bodies within proliferated fibroblasts. This tumor is reviewed with reference to clinical features, histopathology, etiological considerations, and management.