Recent phenotypic analysis of plasma cells showed that normal plasma cells do express the B-cell lineage specific molecule CD19, but their malignant counterpart (myeloma cells) are CD19-. To clarify the meaning of loss of CD19 antigen on myeloma cells, we first compared the expression of CD19 and Pax-5 genes among B cells, normal plasma cells, myeloma cell lines, and primary myeloma cells, because the Pax-5 gene was reported to encode the transcriptional factor, 6-cell-specific activating protein (BSAP), necessary for CD19 gene expression. Neither CD19 nor Pax-5 mRNA could be detected in those primary myeloma cells and cell lines, whereas normal plasma cells did express both CD19 and Pax-5 mRNA. Furthermore, we could confirm that BSAP-binding activity was not detected in the nuclear extract from CD19- myeloma cell line (KMS-5) but was detected in CD19+ B-cell line (Raji by gel shift assay. We further examined the expression of E2A and Id gene, because E2A and Id are considered to be positive and negative regulators in the expression of Pax-5 gene, respectively. However, no significant differences in the expression of the E2A and Id-2 genes could be observed between myeloma cells and normal plasma cells. Therefore, these data suggest that the altered expression of Pax-5, but not E2A or Id, is responsible for the loss of CD19 expression in human myeloma cells, although the underlying mechanism of the altered Pax-5 gene expression remains to be clarified.

By using two-color phenotypic analysis with fluorescein isothiocyanate-anti-CD38 and phycoerythrin-anti-CD19 antibodies, we found that pre-B cells (CD38+CD19+) significantly decreased depending on the number of plasma cells (CD38++CD19+) in the bone marrow (BM) in the cases with BM plasmacytosis, such as myelomas and even polyclonal gammopathy. To clarify how plasma cells suppress survival of pre-B cells, we examined the effect of plasma cells on the survival of pre-B cells with or without BM-derived stromal cells in vitro. Pre-B cells alone rapidly entered apoptosis, but interleukin-7 (IL-7), a BM stromal cell line (KM-102). or culture supernatants of KM-102 cells could support pre-B cell survival. On the other hand, inhibitory factors such as transforming growth factor-pl (TGF-β1) and macrophage inflammatory protein-1p (MIP-1β) could suppress survival of pre-B cells even in the presence of 11-7. Plasma cells alone could not suppress survival of pre-B cells in the presence of IL-7, but coculture of plasma cells with KM-102 cells or primary BM stromal cells induced apoptosis of pre-B cells. Supernatants of coculture with KM-102 and myeloma cell lines (KMS-5) also could suppress survival of pre-B cells. Furthermore, we examined the expression of IL-7, TGF- β 1, and MIPl β mRNA in KM-102 cells and primary stromal cells cocultured with myeloma cell lines (KMS-5). In these cells, IL-7 mRNA was downregulated, but the expression of TGF-p1 and MIP-lp mRNA was augmented. Therefore, these results suggest that BM-derived stromal cells attached to plasma (myeloma) cells were modulated t o secrete lesser levels of supporting factor (IL-7) and higher levels of inhibitory factors (TGF- βI and MIP-1 β) for pre-B cell survival, which could explain why the increased number of plasma (myeloma) cells induced suppression of pre-B cells in the BM. This phenomenon may represent a feedback loop between pre-B cells and plasma cells via BM stromal cells in the BM.

In the peripheral blood (PB) we detected so-called early plasma cells that might already be committed to entering the bone marrow (BM). By two-colour staining with FITC-anti-CD38 antibody, their intensity (CD38++) of expression of CD38 antigen was between that of germinal centre (GC) B cells (low expression (CD38+)) and that of BM plasma cells (high expression (CD38++)), and their phenotype was CD38++ CD19+ CD10- CD20- CD21+ CD24- CD39+ CD5- VLA-4+ VLA-5- MPC-1- without expression of surface membrane IgM (SmIgM). Morphological and immunological examination of the sorted cells confirmed that they were plasmacytoid cells with expression of cytoplasmic IgG (cIgG). Variations of these early plasma cells were examined in various diseases. In active systemic lupus erythematosus, bacterial septicaemia and liver cirrhosis, early plasma cell levels were significantly increased in PB, and after subsidence of such inflammation (inactive states) these cells returned to normal levels. In contrast, normal early plasma cells were significantly suppressed in myelomas, whilst normal or slightly increased numbers of early plasma cells was found in benign monoclonal gammopathy (BMG). In addition, the number of normal early plasma cells returned to a normal level in myeloma cases with complete responses. Therefore, early plasma cells were identified phenotypically, and an increase and decrease in these cells in PB may reflect mobilization and suppression, respectively, of activated B cells into BM plasma cells.

Using two-colour phenotypic analysis with anti-CD38 antibody, human myeloma cells can be classified into VLA-5- immature and VLA-5+ mature cells. We examined the relationship between variations of these subpopulations and clinical responses during treatment in multiple myeloma (MM). 39 patients with MM were treated with combined chemotherapy. First estimation of clinical responses after induction therapy showed that early clinical responses were correlated with the percentage of immature myeloma cells present after induction therapy (P < 0.01), not at diagnosis. After three courses of cyclic maintenance therapy, immature myeloma cells significantly decreased in proportion along with a decrease in total myeloma cells in maintained or more responsive cases (P < 0.01). On the other hand, immature myeloma cells were still found in high proportions in nonresponsive cases with no change (NC) or minor response (MR) (P < 0.01). Furthermore, in relapsing cases from partial response (PR) or progressive disease (PD) from nonresponsive cases, immature myeloma cells increased markedly. Therefore these results show that high proportions of VLA-5- immature myeloma cells remaining after induction therapy and during maintenance therapy correlate well with a declining clinical course of MM during maintenance therapy.

The mature myeloma cells express very late antigen 5 (VLA-5) and MPC-1 antigens on their surface and adhere to bone marrow (BM) stromal cells more tightly than the VLA-5^-MPC-1^- immature myeloma cells in vitro. The VLA-5 and MPC-1 antigens possibly function as two of the molecules responsible for interaction of mature myeloma cells with BM stromal cells. However, the immature myeloma cells do interact with BM stromal cells, and it is unclear which adhesion molecules mediate their interaction. In this study, we found that both immature and mature myeloma cells expressed CD21, an adhesion molecule known to bind to CD23. CD21 was also detected on normal plasma cells. To evaluate the role of CD21 expression on myeloma cells, two myeloma cell lines, NOP-2 (VLA-5-MPC-l-) and KMS-5 (VLA-5'MPC-l'), were used as representatives of immature and mature myeloma cell types, respectively, and an adhesion assay was performed between the myeloma cell lines and BM stromal cells. Antibody-blocking results showed that adhesion of the mature type KMS-5 to KM102, a human BM derived stromal cell line, or to short-term cultured BM primary stromal cells was inhibited by monoclonal antibodies (MoAbs) against CD21, VLA-5, and MPC-1, and inhibition of adhesion of the immature type NOP-2 t o KM102 by the anti-CD21 MoAb was observed as well. Furthermore, CD23 was detected on KM102. Treatment of KM102 with an anti-CD23 MoAb also inhibited adhesion of either KMS-5 or NOP-2 to KM102. Therefore, we propose that CD21 expressed on myeloma cells likely functions as a molecule responsible for the interaction of immature myeloma cells as well as mature myeloma cells with BM stromal cells, and CD23 may be the ligand on the stroma cells for the CD21-mediated adhesion.

Several biomarkers have been checked and verified for use in improving risk rating and management resolutions in cases of community-acquired pneumonia (CAP) alongside the well-established severity scores.
Objective Our objective was to evaluate the veracity of the serum total cortisol level as a biomarker for assessing the severity of CAP.
Patients and methods Our research was a descriptive and prospective study of patients who have been diagnosed with CAP. All were admitted in the Chest Department of Assiut University Hospital between October 2017 and January 2019. The CAP severity was assessed for all enrolled patients by application of the pneumonia severity index (PSI). Serum total cortisol level was measured once in the morning (between 7 a.m. and 8 a.m.) within the first 24 h after admission. All of the patients were followed up until being discharged. The outcome variable was the intrahospital mortality.
Results This research enrolled 94 patients with CAP; of them, 62 (56%) patients survived, whereas the health of 32 (44%) patients deteriorated and they died. The nonsurvivors had significantly higher cortisol level in the serum compared with the survivors (34.15±9.78 vs. 24.90±8.89; P=0.04). Of the study population, 37 (39.4%) patients had normal cortisol level, whereas 57 (60.6%) had high cortisol level. It was noticed that patients with high cortisol level had a significantly higher PSI (140.75±48.31 vs. 103.56±43.57), and 87.2% of them had PSI class V. The serum cortisol level had a positive significant correlation with PSI points (r=0.45, P=0.01). In terms of the diagnostic performance of serum cortisol, it was noticed that serum cortisol at cutoff point more than 43 µg/dl had 80% sensitivity and 91% specificity to predict the intrahospital mortality in patients with CAP.
Conclusion The serum cortisol could be considered as a promising biomarker that is able to predict the severity and outcome of CAP. However, we recommend further larger studies with different aspects to investigate the accuracy of serum cortisol in assessing the severity of CAP and its potential effect on the treatment plan.

Prognostic markers play an essential role in the proper management of community-acquired pneumonia. This research work aimed to evaluate the association of RDW and /or MPV with mortality and morbidity in patients with CAP to improve the yield of already used prognostic scores.

Results

The current study enrolled 153 patients with community-acquired pneumonia (CAP). Out of them, 101 (64%) patients improved while 52 (36%) died. It was noticed that each of delta MPV and RDW (P < 0.001) had positive significant correlation with PSI and CURB-65. Delta MPV and RDW was significantly higher in patients who died (2.61 ± 1.01 vs. 1.78 ± 0.76; P = 0.01 for delta MPV and 16.50 ± 3.54 vs. 15.50 ± 2.81; P = 0.02 for RDW).

Conclusion

Initial RDW and rising MPV during hospitalization for CAP is associated with more severe clinical characteristics and high mortality. Moreover, the use of RDW and delta MPV in patients admitted with CAP can improve the performance of prognostic scales.

The frequency of fungal infections of the lung has increased particularly in immunocompromised pa-tients. Early diagnosis and treatment is important to start antifungal therapy and to avoid unnecessary use of toxic antifungal agents. This study aimed to identify the common fungal species causing pulmonary infection in immunocom-promised patients and their in vitro antifungal sensitivity pattern in Assiut University Hospitals (AUH).
Subjects and Methods: This was a hospital based descrip-tive study conducted on 135 patients admitted at different Intensive Care Units (ICUs) and Oncology Department at Assiut University Hospitals (AUH). Collected respiratory specimens were subjected to direct microscopic examination and inoculation on Sabouraud Dextrose Agar (SDA). Identi-fication of isolated yeasts was done using phenotypic methods including chromogenic media (Brilliance Candida agar and CHROMagar Candida differential media), germ tube test, cornmeal agar and API candida while mould isolates identifi-cation was mainly dependent on macroscopic and microscopic features. Some isolates had confirmed using rRNA gene sequencing. In vitro antifungal susceptibility testing was done using disc diffusion method.
Results: In this study 80/135 (59.3%) of collected samples were positive for fungal infection. The most common fungal pathogens isolated were Candida and Aspergillus species. In vitro sensitivity test showed that the yeast isolates had the highest sensitivity to Nystatin (90.9%) followed by Ampho-tericin B (80.3%) while for mould isolates, the highest sensi-tivity was to Voriconazole (71.4%) followed by Amphotericin B (57.1%).
Conclusion: Pulmonary fungal infection appears to be an important problem in immunocomprimised patients with Candida albicans was the most commonly isolated yeast from various clinical specimens; also the increase in the resistance especially to azoles is a major concern.

Aim of the work: To investigate the association of single nucleotide polymorphism (SNP) (rs2073618) of the OPG gene and of serum OPG with subclinical carotid atherosclerosis in RA patients. Patients and methods: Eighty RA patients with no previous history of a cardiovascular disease were studied and forty healthy controls were enrolled in the study. Carotid atherosclerosis was evaluated by highresolution B-mode ultrasound and the carotid intimal medial thickness (CIMT) measured. rs2073618 OPG genotyping was performed by polymerized chain reaction (PCR) and serum OPG concentrations were measured. The high sensitive C reactive protein (hs-CRP), rheumatoid factor (RF) titer and anti-cyclic citrullinated peptide (anti-CCP) were assessed. The disease activity score (DAS28) was evaluated. Results: The patients mean age was54.1 ± 6.2 years, disease duration of 12.5 ± 8.5 years and were 72 females and 8 males. Increased IMT was found in 38 patients, 40 age and sex matched controls were included. Patients with atherosclerosis (n = 38) had longer disease duration, higherDAS28, hs-CRP, RF titer and anti-CCP. The serum OPG levels were higher in patients with atherosclerosis (1106.4 ± 1157.1 ng/l) compared to those without (658.3 ± 151.1 ng/l)(p = 0.001). Serum OPG significantly correlated with disease duration (r = 0.42, p = 0.005), DAS28 (r = 0.53, p = 0.001), hs-CRP (r = 0.41, p = 0.007), anti-CCP (r = 0.47, p = 0.003) and mean CIMT (r = 0.37, p = 0.02). The frequencies of CC, CG and GG genotypes were comparable between those with and without atherosclerosis (39.5%, 50%, 10.4% vs 42.9%, 47.6% and 9.5% respectively). Conclusion: rs2073618 OPG gene may not be associated with subclinical atherosclerosis, although the serum level could be a reliable marker for disease activity and for early detection of carotid artery atherosclerosis in RA.