Cyclophosphamide

Basic descriptive statistics (mean, median, standard deviation, interquartile range, skewness, and kurtosis) were computed for all variables, which were subsequently tested for normality using the Kolmogorov–Smirnov and Shapiro–Wilk tests. Differences in interval variables (e.g. Hsp90, age, etc.) were tested using the Mann–Whitney U test, while the chi-square test was used to compare frequency counts of categorical variables (e.g. gender). The bivariate relationships between variables under study were assessed using the Spearman correlation coefficient. Linear regression analysis was used to predict patients' Hsp90 levels by a set of predictors (FEV1, FVC, DLCO, SpO₂) while adjusting for a confounder (CRP). Friedman's test was used to analyze repeated longitudinal measurements taken: (a) at baseline, (b) after 1 month, (c) after 6 months, and (d) after 12 months of therapy with cyclophosphamide. Data are presented as median (IQR) unless stated otherwise. Statistical significance was set at p < 0.05. All analyses were conducted using SPSS version 25 (SPSS, Inc., Chicago, IL, USA). Graphs were created using GraphPad Prism 5 (version 5.02; GraphPad Software, La Jolla, CA, USA).

All patients involved in this study fulfilled the ACR/EULAR classification criteria for SSc^{43 (link)}. Plasma samples from all patients and healthy individuals were prepared from the whole blood collected into commercially available EDTA-treated tubes. For the cross-sectional analysis, plasma samples were obtained from 92 Caucasian patients with systemic sclerosis (SSc) and 92 age- and sex-matched Caucasian healthy controls. For the longitudinal analysis, plasma samples were obtained prospectively at baseline, and 1, 6, and 12 months thereafter from 30 Caucasian patients with SSc-ILD with active alveolitis without pulmonary arterial hypertension who underwent a routine 6-month (n = 16) or 12-month (n = 14) treatment with i.v. cyclophosphamide (CPA, 500 mg/m² monthly). Active alveolitis was defined as the presence of areas of ground-glass attenuation on high-resolution computed tomography (HRCT) and reduced levels of diffusing lung capacity for carbon monoxide (DLCO) and/or forced vital capacity (FVC), as described elsewhere^{44 (link)}. Other SSc-related clinical features were assessed according to generally accepted definitions and recorded, such as the presence of pulmonary arterial hypertension, renal, cardiac and gastrointestinal involvement, Raynaud's phenomenon, and digital ulcers^{45 (link)}. Skin involvement was evaluated using the modified Rodnan skin score (mRSS)^{46 (link)}. Disease activity was determined by the European Scleroderma Study Group (ESSG) SSc activity score^{47 (link)}. Pulmonary function tests (PFT) were routinely performed using standard methods, in accordance with the ATS recommendations^{48 (link)}. The DLCO was measured by a single-breath method using a gas mixture of 0.2% CO and 8% helium, with correction for hemoglobin. Peripheral oxygen saturation (SpO₂) was measured by a handheld pulse oximeter (CR-100, Noramedica, Czech Republic). In the longitudinal analysis PFTs were performed at baseline, and 6 and 12 months thereafter, and the results are expressed as a percentage of the normal predicted values based on the patient’s sex, age, and height. The research was confirmed by the local ethics committee at the Institute of Rheumatology in Prague, and each patient signed an informed consent form. All methods were performed in accordance with the relevant guidelines and regulations.

We included all breast cancer patients that were operated on at the Karolinska Hospital from 1 January 1994 to 31 December 1996 (n = 524), identified from the population-based Stockholm–Gotland breast cancer registry established in 1976. Available tumor material was frozen on dry ice or in liquid nitrogen and was stored in -70°C freezers. Figure 1 shows the details of various exclusions leading to the final 159 patients for analysis. The ethical committee at the Karolinska Hospital approved this microarray expression project.
The different reasons for exclusion were not influenced by age at diagnosis (Table 1). The 231 tumors that were not analyzed using expression profiling had a lower mean diameter, had fewer mean affected lymph nodes, and had fewer individuals with recurrent disease at the end of the study period (Table 1). For those excluded for other reasons, there did not seem to be a selection based on age or stage of the disease, compared with those patients included in the study (Table 1).
The Stockholm–Gotland Breast Cancer Registry, supplemented with patient records, were examined for information on the tumor size, the number of retrieved and metastatic axillary lymph nodes, the hormonal receptor status, distant metastases, the site and date of relapse, initial therapy, therapy for possible recurrences, the date and cause of death. Tumor sections from the primary tumors from patients with array profiles were classified using Elston–Ellis grading [18 (link)] by a blinded pathologist (HN).
In the adjuvant setting tamoxifen and/or goserelin is normally used for hormonal treatment, but mostly intravenous cyclophosphamide, methotrexate and 5-fluorouracil (CMF) on days 1 and 8 was used as adjuvant chemotherapy, except in high-risk patients who were offered inclusion in the Scandinavian Breast Group 9401 study [19 (link)]. After primary therapy, patients were recommended to have regular clinical examinations and yearly mammograms, in addition to laboratory and X-ray tests guided by clinical signs and symptoms. Patients were normally followed for 5 years. Patients followed up outside the Karolinska Hospital were tracked using a unique personal identification number. There was no loss to follow-up.
The relapse site, date of relapse, relapse therapy and date of death were ascertained in May 2002. The average follow-up was 6.1 years. Cause of death was coded as death due to breast cancer (including those with distant metastases but dying from other causes), death due to other malignancies and death due to nonmalignant disorders. Through the population-based Swedish Cancer Registry, second primary malignancies were identified.

ANDROMEDA is a randomized, open-label, active-controlled, phase 3 study. Patients were randomized (1:1) to receive VCd with or without DARA SC (daratumumab 1800 mg co-formulated with recombinant human hyaluronidase PH20 [2000 U/mL; ENHANZE^® drug delivery technology, Halozyme, Inc., San Diego, CA, USA]). All patients received bortezomib 1.3 mg/m² subcutaneously, cyclophosphamide 300 mg/m² orally or intravenously [500 mg maximum weekly dose], and dexamethasone 40 mg orally or intravenously once weekly for 6 cycles of 28 days each. DARA SC was administered by manual injection over approximately 5 min weekly in cycles 1 and 2, every 2 weeks in cycles 3–6, and every 4 weeks thereafter until disease progression, until the start of subsequent therapy, or for a maximum of 24 cycles from the start of the study, whichever occurred first. The median follow-up period was 11.4 months. See Online Resource 1 (Supplementary Methods) for additional details.

The primary therapeutic modalities were determined using the Lugano and Paris staging system (Online Resource 1) and the HPI status. H. pylori eradication was performed in all patients with HPI and localized stage gastric MALT lymphoma. For first-line eradication therapy, a proton pump inhibitor (PPI)-based triple therapy regimen was administered for 2 weeks: PPI (standard dose twice a day), clarithromycin (0.5 g twice a day), and amoxicillin (1 g twice a day). 13C urea breath tests were performed in all patients for 3 months or at least 8 weeks after treatment completion, and at least 2 weeks after PPI withdrawal to confirm HPI eradication. For patients who failed first-line triple therapy, a second-line quadruple-therapy regimen consisting of PPI (standard dose twice a day), tripotassium dicitrato bismuthate (300 mg four times a day), metronidazole (500 mg thrice a day), and tetracycline (500 mg four times a day) was administered for 1–2 weeks.
Patients received radiotherapy, chemotherapy, or chemoradiotherapy if they did not achieve lymphoma regression following first- and second-line HPI eradication therapy, or were at the localized stage without initial HPI, or had advanced-stage gastric MALT lymphoma. For radiotherapy, the clinical target volume included the entire stomach and regional lymph nodes and was prescribed as 30.6 Gy over 17 fractions on the stomach [20 (link)]. The internal target volume (ITV) and planning target volume were set using the motion information obtained from the 4-dimensional CT for assessment of breathing motions and defined as an expansion of 5 mm from the ITV considering the set-up error of the patient [20 (link)]. Patients with the involvement of ≥ 2 organs were excluded from radiotherapy. The R-CVP was the primary systemic chemotherapy regimen, consisting of rituximab 375 mg/m², cyclophosphamide 750 mg/m², and vincristine 1.4 mg/m² on day 1, and prednisolone 60 mg/m² on days 1–5 every 21 days. Localized stage lesions involving small-sized mucosal layers in patients with initial HPI-negative findings could be selectively treated by endoscopic mucosal resection (EMR) and close observation. In the case of chemoradiotherapy, we only used additional radiotherapy for consolidation purposes after chemotherapy by the physicians’ decision. To investigate the side effects of each treatment modality, we reviewed the medical records following the National Cancer Institute’s Common Terminology Criteria for Adverse Events version 5.0.