[Concurrent COVID-19 and Pneumocystis Jirovecii Pneumonia in a Patient with Breast Cancer Receiving Adjuvant Dose-Dense Chemotherapy].

A 49-year-old woman had been treated with adjuvant dose-dense chemotherapy for left breast cancer pT2N2aM0, Stage ⅢA. She developed a fever of over 38°C on day 13 of the third course of dose-dense doxorubicin/cyclophosphamide (AC)chemotherapy. She was started on oral levofloxacin, but the fever did not resolve. COVID-19 PCR test was positive and chest CT scan showed bilateral ground-glass opacities. She was diagnosed with COVID-19 pneumonia and hospitalized. The fever did not resolve even after sotrovimab and remdesivir were administered. On the 5th hospital day, the serum ß-D-glucan level was found to be elevated(81.7 pg/mL), and she was diagnosed with concurrent COVID-19 and Pneumocystis jirovecii pneumonia(PCP). After the start of sulfamethoxazole-trimethoprim(TMP-SMX), the fever resolved quickly. After discharge from hospital, ground-glass opacities had disappeared. She resumed dose-dense chemotherapy with TMP-SMX and completed without fever. In immunosuppressed patients with cancer drug therapy, it is necessary to make a differential diagnosis of various types of pneumonia and to consider the different types of pneumonia may occur simultaneously.

Landmark Analysis Exploring the Optimal Period for Intensive Monitoring After Return to Work for Employees With Sickness Absence Due to Common Mental Disorders.

OBJECTIVE: This study aimed to clarify when and how long intensive monitoring should be performed after return to work (RTW) of employees experiencing common mental disorders using landmark analysis. METHODS: We conducted a retrospective survey of workers who experienced sickness absences (SAs) during 36 months after RTW. Sustainability rates of attendance among the following groups were compared before and after the landmark (18 months): one SA episode (Group 1), two SA episodes (Group 2), and three or more SA episodes (Group 3). RESULTS: Before the landmark, sustainability in Group 1 was higher than in the other groups, with no significant differences among groups after the landmark. Sustainability rate of attendance tended to be lower before than after the landmark in each group. CONCLUSIONS: Intensive monitoring is suggested in the first 18 months after RTW.

Number of Previous Absences Is a Predictor of Sustained Attendance After Return-to-Work in Workers With Absence due to Common Mental Disorders: A Cohort 3-Year Study.

OBJECTIVE: This study evaluated the relationship of the number of previous episodes due to common mental disorders (CMDs) with long-term outcomes and sustainability of attendance after return-to-work (RTW). METHODS: Participants were assigned to the following three groups: workers having one (Group 1), two (Group 2), and three or more (Group 3) previous episodes. Outcomes were a recurrent absence and the sustainability rate of attendance after RTW. RESULTS: The sustainability rate in Group 1 was significantly higher than that in Group 3 throughout the observation period. The sustainability rates for Group 2 were significantly higher than for Group 3 at 30 and 36 months. CONCLUSIONS: The number of previous episodes was shown to affect sustainability of attendance after RTW due to CMDs, indicating that repeated previous absences are a significant prognostic factor.

LATS2 promoter hypermethylation and its effect on gene expression in human breast cancer.

Tumor-specific promoter hypermethylation of large tumor suppressor, homolog 2 (LATS2), a tumor suppressor gene, has been investigated using methylation-specific polymerase chain reaction (MSP) assays in different types of human cancer producing conflicting results. The aim of the present study was to evaluate the methylation status of the LATS2 promoter region using bisulfite sequencing with a next generation sequencer for breast cancer. In the 11 patients enrolled in the present study, the LATS2 promoter methylation index (MI) was uniformly high in tumor and normal tissues of the breast (median, 84.0 and 87.4%, respectively). The presence of LATS2 promoter hypermethylation was confirmed in isolated tumor cells and normal epithelial cells using the magnetic-activated cell sorting method. In situ hybridization for LATS2 messenger RNA (mRNA) revealed that the mRNA expression of LATS2 was higher in normal epithelial cells, compared with tumor cells, however, it was not significantly associated with LATS2 MI. In 12 breast cancer cell (BCC) lines and two normal breast cell lines, the LATS2 promoter was uniformly hypermethylated with no correlation between the mRNA expression of LATS2 and the LATS2 MI. In addition, treatment of the BCC lines with a demethylating reagent had minimal effect on the mRNA expression of LATS2 in any of these cell lines. These results demonstrated that LATS2 hypermethylation was not involved in silencing the mRNA expression of LATS2 mRNA. The lower mRNA expression level of LATS2 in tumor cells, compared with normal epithelial cells, suggested the possible involvement of downregulation in the mRNA expression of LATS2 in the pathogenesis of breast cancer. Therefore, the conflicting results previously reported for LATS2 promoter methylation in different types of cancer, detected using MSP assays may be attributable to the low fidelity of the MSP assay.

Mutational Analysis of AKT1 and PIK3CA in Intraductal Papillomas of the Breast with Special Reference to Cellular Components.

The pathologic feature of intraductal papillomas is defined as a papillary structure composed of a fibrovascular stromal core lined by luminal epithelial cells and myoepithelial cells. We used droplet digital PCR for the mutational analysis of AKT1 (E17K) and PIK3CA (H1047R, E542K, and E545K) in 60 papillomas. AKT1 and PIK3CA mutations were detected in 12 (20%) and 17 (28%) of the papillomas, respectively. In five tumors harboring mutations, mutational analysis of AKT1 or PIK3CA was performed separately using luminal epithelial cells and myoepithelial cells sorted using anti-cytokeratin 19 antibody and anti-α smooth muscle actin antibody. The two types of cells from a given papilloma had the identical mutation. Three patients with the PIK3CA mutation-positive papilloma developed breast cancers at the resection site of the papilloma, but none of these subsequent breast cancers had the PIK3CA mutation. These results indicate that a papilloma stems from a bipotent progenitor cell that contains the AKT1 or PIK3CA mutation and proliferates and differentiates to form the papilloma. Papilloma can be a risk factor for developing breast cancer but is unlikely to be its obligate precursor.

Methylation of the SEPT9_v2 promoter as a novel marker for the detection of circulating tumor DNA in breast cancer patients.

The aim of the present study was to evaluate the promoter methylation status of SEPT9_v2 in breast cancer and to detect this methylated gene in circulating tumor DNA (ctDNA) in plasma. Bisulfite sequencing was performed with a next generation sequencer. Methylation of the SEPT9_v2 promoter was found in 67% (8/12) of breast cancer cell lines and 53% (10/19) of breast tumor tissue, but not in normal breast tissue (0/19). A clear inverse correlation was observed between the expression of SEPT9_v2 mRNA and the methylation index (MI) both in cell lines and breast cancer tissues. The MI of SEPT9_v2 was significantly higher in non-basal subtype of breast cancer (13.0%, n=84) than in basal subtype (3.0%, n=23) (P<0.0001). Methylated SEPT9_v2 ctDNA in plasma was detected in 11% (9/82) of primary breast cancer patients and 52% (26/50) of metastatic breast cancer patients, but not in the healthy controls (0/51). These results indicate that SEPT9_v2 promoter hypermethylation, which silences the expression of SEPT9_v2 mRNA, is observed in a significant proportion of breast tumors, and that methylated SEPT9_v2 may serve as a novel tumor marker for breast cancer.

Loss of imprinting of IGF2 in fibroadenomas and phyllodes tumors of the breast.

Loss of imprinting (LOI) of insulin-like growth factor 2 (IGF2) is thought to be implicated in the pathogenesis of some tumors by upregulating IGF2 mRNA but its role in the pathogenesis of fibroadenomas (FAs) and phyllodes tumors (PTs) of the breast is yet to be studied. LOI of IGF2 was investigated in 25 FAs and 17 PTs which were heterozygous for Apa I polymorphism, and was found to be present in 13 FAs and 12 PTs. IGF2 mRNA expression was more upregulated in FAs and PTs than in paired surrounding normal tissues and laser microdissection showed that IGF2 mRNA expression was significantly higher in the stromal than the epithelial cells. LOI was not associated with upregulation of IGF2 mRNA, nor were MED12 mutations and methylation status of the differentially methylated region 0 (DMR0) of IGF2. These results demonstrate that IGF2 mRNA expression is more upregulated in FAs and PTs than in normal tissues, especially in their stromal cells, but such an upregulation is not related to LOI of IGF2, and that hypomethylation of DMR0 is unlikely to be involved in induction of LOI.

Promoter methylation of TRIM9 as a marker for detection of circulating tumor DNA in breast cancer patients.

The aim of the present study was to investigate the promoter methylation status of TRIM9 in breast cancer and to determine the presence of TRIM9-methylated circulating tumor DNA (ctDNA) in plasma. Bisulfite sequencing with a next generation sequencer showed TRIM9 promoter methylation in 92 % (11/12) of breast cancer cell lines (BCCs) and 68 % (13/19) of breast tumor tissues but not in any normal breast tissues (0/19). Methylation ratio of TRIM9 was significantly lower in basal type (9 %, n = 23) than luminal A (69 %, n = 29, P = 0.0003). Quantitative RT-PCR of BCCs disclosed an inverse correlation between TRIM9 mRNA expression and methylation ratio. TRIM9 methylated ctDNA in plasma was detected in 18 % (10/56) of metastatic breast cancer patients but not in any of 60 healthy controls. These results indicate that TRIM9 promoter hypermethylation, which suppresses TRIM9 mRNA expression, occurs in a significant proportion of breast tumors, and that TRIM9-methylated ctDNA thus may serve as a tumor marker for breast cancer.

Mutational analysis of MED12 in fibroadenomas and phyllodes tumors of the breast by means of targeted next-generation sequencing.

We aimed to analyze MED12 mutation in fibroadenomas (FAs) and phyllodes tumors (PTs) of the breast, which are closely related and consist of epithelial and stromal components. Targeted deep-sequencing using next-generation sequencing was performed in FAs (n = 58) and PTs (n = 27). The frequency of MED12 mutant tumors was significantly higher (P = 0.016) in PTs (74.1 %) than in FAs. (46.6 %). As for FAs, this frequency was significantly higher (P = 0.001) for intracanalicular type (69.0 %) than for other histological subtypes such as pericanalicular, organoid, and mastopathic types (24.1 %). Laser microdissection study revealed that stromal cells, but not epithelial cells, harbored MED12 mutations in both FAs and PTs. MED12 mutation is implicated in the pathogenesis of both FAs and PTs. The similarly high frequency of MED12 mutation in intracanalicular type FAs suggests that they are most closely related to PTs. It is thus speculated that FAs with MED12 mutation are more likely to progress to PTs.