Few studies have examined variations in both the physical signs and clinical findings in the final days and hours before the death of elderly patients. We determined the physical signs and clinical findings in patients who were at least 75 years old or were diagnosed with cancer with impending death and examined the association of these parameters with the profile and timing of their endocardiography (ECG) and oxygen saturation (SpO) changes prior to death. In this prospective, observational study, between April 2014 and June 2017, we enrolled elderly patients who were admitted to the Respiratory Medicine Ward in our hospital and were near death, which was determined based on certain symptoms such as a loss of oral intake. We recorded their physical signs (oral intake, consciousness level, and respiration with mandibular movement) 4 times a day from admission to death. We evaluated their changes in ECG and in SpO levels for up to 24 hours preceding death. For the 70 patients who died in our ward, we found a loss of oral intake at 6 days, consciousness impairment at 1.3 days, and then respiration with mandibular movement at 12 hours before death were the most consistent findings before death. When we analyzed the ECG and SpO changes during impending death, 83% of the patients showed undetectable SpO levels followed by a loss of heart rate. The loss of P wave in the ECG was characterized as process on impending death. Respiration with mandibular movement was one of the specific signs of impending death in ill elderly patients. The monitoring of ECG and SpO levels may be a useful tool to predict the impending time of death.

Mutations in the epidermal growth factor receptor (EGFR) gene are associated with a favorable clinical response to the EGFR tyrosine kinase inhibitors gefitinib and erlotinib in non-small cell lung cancer (NSCLC). We present here, a new method for the rapid detection of the two most common EGFR mutations (delE746-A750 and L858R) from clinical samples. The methodology involves the combination of newly designed mutation-specific primers and a novel real-time PCR machine with an innovative thermo-control mechanism that enables ultrarapid PCR. We evaluated this method using a cell mixture composed of various ratios of lung cancer cells harboring mutated or wild-type EGFR, lung cancer tissues obtained by surgery, and a cytology sample obtained by bronchoscopy from a lung cancer patient. In the cell mixture analysis, our method detected 0.1% of cells with delE746-A750 and 1% of cells with L858R among cells with wild-type EGFR. In 143 lung cancer tissues, the result of this assay was concordant with those of direct sequencing in 138 samples. The five samples with discordant results were tested using a PCR-Invader assay and the result matched those of our method at 100%. We also successfully detected EGFR mutations in the lavage obtained from a lung cancer patient. The turnaround time for this method was <10 min, and all steps could be accomplished in <50 min after sample collection. Thus, our novel PCR method offers a rapid, simple, and less expensive test for EGFR mutations and can be applied as a point-of-care diagnostic test.

Abstract. Small cell lung cancer (SCLC) is one of the intractable malignancies. The goal of this study was to clarify whether Akt activity is involved with chemo-resistance and to improve the sensitivity of SCLC cells to the current standard chemotherapeutic drugs with agents that are expected to suppress Akt activity through tyrosine kinase inhibition. Although Akt activity seemed to be involved with the sensitivity of SCLC cells to chemotherapeutic agents (cisplatin, etoposide, SN38 and amrubicin), in Akt-activated N417 cells, only amrubicin exerted synergistic cell growth inhibition when combined with an Akt inhibitor, LY294002. A non-specific tyrosine kinase inhibitor, genistein, suppressed Akt and showed synergistic interaction in combination with amrubicin in N417 cells. Among tyrosine kinases (insulin-like growth factor I receptor, c-Kit and c-Src), only c-Src was activated in N417 cells compared with Akt-inactive H209 cells. A c-Srcspecific inhibitor, PP2, and a clinically available multi-tyrosine kinase inhibitor, dasatinib, suppressed Akt activity in parallel with c-Src inhibition. Both PP2 and dasatinib exerted synergistic growth inhibition of N417 cells in the combination with amrubicin. In immunohistochemical analysis, c-Src was expressed in 17 of 19 of the SCLC tumor tissues. These observations suggested that Akt suppression enhances the cytotoxicity of amrubicin, and for the purpose of Akt suppression, c-Src is a promising target in SCLC.

An epidermal growth factor receptor (EGFR) mutation is the best marker of sensitivity to the EGFR tyrosine kinase inhibitors gefitinib and erlotinib. Polymerase chain reaction (PCR)-based methods combined with the use of fluorescence probes, such as the Scorpion Amplification Refractory Mutation System (ARMS) or the PCR-Invader method, are frequently used to detect EGFR mutations before therapy. The sensitivities and specificities of these methods are satisfactory; however, there is potential to further improve the cost and detection time. We developed a new method for the rapid and reliable detection of EGFR mutations by using a combination of mutation-specific primers and the newly developed ultra-rapid real-time PCR (Hyper-PCR) amplification method that can be accomplished in &lt;7 min. We designed specific ARMS primers for the common EGFR del E746-A750 mutations, and optimized the reaction conditions for the PCR machine that contained a thin disc-type reaction vessel, the temperature of which could be quickly altered by rotating it on thermal sources. We used Hyper-PCR to analyze the common EGFR mutations del E746-A750 and L858R in lung cancer samples from 143 non-small cell lung cancer (NSCLC) patients. The results were compared with those obtained by direct sequencing, conventional PCR, and the PCR-Invader method. Using the direct sequencing method, 13 (9.1%) and 15 (10.5%) NSCLC samples were positive for the del E746-A750 and L858R mutations, respectively, while the Hyper-PCR method revealed that 15 (10.5%) and 18 (12.6%) samples were positive for the del E746-A750 and L858R mutations, respectively. Five samples, whose results were discordant between the Hyper-PCR and direct sequencing methods, were analyzed by conventional PCR and the PCR-Invader method, and the results were found to be consistent with those obtained by Hyper-PCR. These results suggest that the novel Hyper-PCR method is an inexpensive, rapid, and reliable diagnostic test for EGFR mutations. Citation Format: Hiroki Chikumi, Miyako Takata, Keiji Matsunami, Shingo Matsumoto, Masahiro Kotani, Tomohiro Sakamoto, Hirokazu Touge, Kosuke Yamaguchi, Jun Kurai, Naomi Miyake, Masaki Nakamoto, Tadashi Igishi, Eiji Shimizu. A new rapid diagnostic test to identify epidermal growth factor receptor mutations in non-small cell lung cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-88. doi:10.1158/1538-7445.AM2013-LB-88

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