Additional statin treatment enhances the efficacy of HER2 blockade and improves prognosis in Rac1-high/HER2-positive breast cancer.

The prognosis of HER2-positive breast cancer (BC) has improved with the development of anti-HER2 therapies; however, the problem remains that there are still cases where anti-HER2 therapies do not respond well. We found that the expression of SREBF2, a master transcriptional factor in the mevalonate pathway, was correlated with ERBB2 (HER2) expression and a poor prognosis in HER2-positive BC. The target gene expressions of SREBF2 were associated with higher expression of ERBB2 in HER2-positive BC cells. Statins, anti-hypercholesterolemia drugs that inhibit the mevalonate pathway, enhanced the efficacy of HER2-targeting agents with inducing apoptosis in a geranylgeranylation-dependent manner. Mechanistically, statins specifically inhibited membrane localization of Rac1, a target protein of geranylgeranylation, and suppressed the activation of HER2 downstreams AKT and ERK pathways. Consistently, retrospective analysis showed a longer recurrence-free survival in Rac1-high/HER2-positive BC patients treated with HER2-targeting agents with statins than without statins. Our findings thus suggest that Rac1 expression could be used as a biomarker to stratify HER2-positive BC patients that could benefit from dual blockade, i.e., targeting HER2 with inhibition of geranylgeranylation of Rac1 using statins, thereby opening avenues for precision medicine in a new subset of Rac1-high/HER2-positive BC.

Validation of late recurrence prediction by gene expression profiles and clinicopathological factors in estrogen receptor-positive breast cancer.

BACKGROUND: The mechanism of late recurrence (LR) of estrogen receptor (ER)-positive breast cancer remains unclear, as previous studies have separately investigated "gene expression profiles" and "clinicopathological factors." Thus, this study aimed to evaluate the predictive capability of LR by combining the two independent factors of gene expression profiles (42-gene classifier: 42GC) and clinicopathological factors (Clinical Treatment Score post-5 years: CTS5) in multiple large cohorts. METHODS: We analyzed microarray CEL file data downloaded from public databases of 28 global cohorts. A total of 2,454 patients with ER-positive breast cancer were analyzed for 42GC, and 1,263 of these, with complete clinicopathological data were analyzed for CTS5. RESULTS: In the analysis of recurrent patients, the 42GC LR and CTS5 low-risk group tended to have LR. Notably, in the analysis of patients with and without recurrence, the highest LR rate beyond 5 years was observed in the CTS5 high-risk group. The combination of the 42GC and CTS5 high-risk groups showed the highest LR rate (16.9%), significantly exceeding that of the 42GC non-LR (NLR) and CTS5 low-risk combination (5.41%) (p = 0.038, odds ratio = 3.53). Furthermore, incorporating a third factor, 95GC, potentially reduced the number of patients prioritized for extended hormonal therapy for approximately one-quarter of patients. CONCLUSIONS: Results confirmed that the two factors, gene expression profiles and clinicopathological factors, affect the time of recurrence. It also showed that the biological predisposition for LR (CTS5 low-risk) differed from the high LR rate (CTS5 high-risk). In clinical practice, patients with the 42GC LR and CTS5 high-risk combination should be prioritized for extended hormonal therapy. The addition of CTS5 and 95GC to 42GC allows for better risk classification of LR.

Investigation of recurrence prediction ability of EndoPredict® using microarray data from fresh frozen tissues in ER-positive, HER2-negative breast cancer and indication expansion of EndoPredict® from microarray data from fresh-frozen to FFPE tissues.

BACKGROUND: EndoPredict® (EP) is a multigene assay to predict distant recurrence risk in luminal breast cancer. EP measures the expression of 12 genes in primary tumor by qRT-PCR from formalin-fixed paraffin-embedded (FFPE) tissues and calculates EP risk score that indicates the risk of distant recurrence. We evaluated the performance of EP in predicting distant recurrence risk using microarray data from fresh frozen (FF) tissues. We also examined the applicability of EP to microarray data from FFPE tissues. METHODS: We analyzed the publicly available data of 431 node-negative and 270 node-positive patients with luminal breast cancer who received endocrine therapy alone. We evaluated the prognostic value of EP using microarray data from FF tissues. Next, we created an algorithm to calculate EP risk score using microarray data from FFPE tissues. We examined the correlation coefficient of EP risk score and concordance rate of EP risk high/low using microarray data from FFPE/FF tissue pairs in a validation set of 39 patients. RESULTS: In 431 node-negative patients, the distant recurrence-free survival (DRFS) rate was significantly worse in those with high EP risk scores (P = 3.68 × 10-6, log-rank). The 5-year DRFS was 95.2% in those with low EP risk score. In the validation set, the correlation coefficient of EP risk score was 0.93 and the concordance rate of EP risk high/low was 91.7%. CONCLUSIONS: EP using microarray data from FF tissues was useful in predicting distant recurrence risk in luminal breast cancer, and EP might be utilized in microarray data from FFPE tissues.

Subjective Symptoms Linked to Sleep Duration: An Analysis from Japanese National Statistics.

BACKGROUND: There is a high prevalence of sleep disorders in Japan, and they are a factor in a decreased quality of life. The main objective of this study was to clarify the background factors of sleep disorders that affect sleep duration, such as subjective symptoms and working hours. METHODS: We performed a cross-sectional study on the Japanese national statistics data. Answers from a household questionnaire were used to analyze risk factors for decreases in sleep duration. The subjects were a total of 3972 men and women aged 40-59 years, the age group that forms the core of the working population. For the analysis, a univariate analysis (contingency table) between sleep duration (two groups: sleep duration ≥ 6 h and <6 h) and 42 subjective symptoms was carried out. A multivariate analysis (binomial logistic regression) was conducted using sleep duration and subjective health assessment as objective variables, and odds ratios (ORs) adjusted for sex, working hours, and other factors were obtained. RESULTS: The univariate analysis by subjective symptom showed significant ORs for eight symptoms, including poor sleep quality (OR: 2.24), constipation (OR: 2.24), and dizziness (OR: 1.77). In the multivariate analysis, the model with sleep duration as the objective variable showed significantly adjusted ORs for four variables, including constipation (1.72) and poor sleep quality (1.66). The model with subjective health assessment as the objective variable showed significantly adjusted ORs for eight variables, including dizziness (4.18), while poor sleep quality (1.45) was not significant. CONCLUSIONS: The present results suggest the presence of subjective symptoms that may be inferred to be related to decreases in sleep duration.

Multi-gene assay 95- and 155-gene classifiers for prognosis prediction and chemotherapy omission in lymphnode positive luminal-type breast cancer.

BACKGROUND: The prognosis of lymphnode positive breast cancer is worse than that of lymph node negative breast cancer but some cases may not require chemotherapy. We investigated the ability of the new multi-gene assays, 95GC and 155GC, to identify patients with lymphnode positive Luminal-type breast cancer whose chemotherapy can be omitted relatively safely. PATIENTS AND METHODS: We extracted 1721 cases of lymphnode positive Luminal-type breast cancer from 22 public database Caucasoid cohorts and 3 Asian cohorts, and performed recurrence prognosis analysis with 95GC and 155GC. RESULTS: Using 95GC, the cases were stratified as the high (n = 917) and low (n = 202) groups according to the prognosis of lymphnode positive Luminal-type endocrine only breast cancer. The 5 years DRFS in the low risk group was relatively good at 90%, and no additional effect of chemotherapy was observed, suggesting omission of chemotherapy. The recurrence prognosis was also significantly dichotomized into the high and low risks by 95GC in 21GC RS 0-25 cases. Here, we found a group with poor prognosis even in post-menopause RS 0-25 and requiring chemotherapy. Additionally, a group in which the prognosis was good in pre-menopause RS 0-25, and the omission of chemotherapy could be considered. Patients in the high-risk group at 155GC had poor prognosis after chemotherapy. 155GC also showed a group that chemotherapy alone was not sufficient. CONCLUSION: In this study, we demonstrated the possibility of accurately selecting patient groups for which chemotherapy can be omitted from lymphnode positive Luminal-type breast cancer.

Optimal Treatment of Hormone Receptor-positive Advanced Breast Cancer Patients With Palbociclib.

BACKGROUND/AIM: Palbociclib was the first cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitor approved worldwide. Currently, CDK4/6 inhibitors are strongly recommended for endocrine therapy in the first or second line with hormone receptor-positive advanced breast cancer. It is expected the use of CDK4/6 inhibitor will further increase. Therefore, the aim was to investigate and better understand the use of palbociclib. PATIENTS AND METHODS: We retrospectively analyzed the data of patients with advanced breast cancer who were treated with palbociclib in three hospitals between 2018 and 2022. Clinical data were obtained from the patients' medical electronic records. RESULTS: A total of 143 patients were enrolled. The median age was 66 years (range=33-89), and the majority (90.9%) were postmenopausal patients. In total, median time-to-treatment discontinuation (TTD) (95% confidence interval, CI) was 7 (6-10) months. Median TTD (95% CI) was 13 (7-20) months for the first or second line, and significantly prolonged compared to TTD for the third or later lines with palbociclib (p<0.0001). The importance of front-line use was indicated. Multivariate analyses showed that no visceral metastasis or first or second line therapy influenced the longer TTD. Between patients above or below 70 years of age, older age did not negatively affect TTD, though there were significantly more cases of dose reduction or withdrawal in patients over 70 years old. The variation of adverse events (AEs) among hospitals was very large (9.0%, 31.3%, 4.5%). We found that understanding of AE management was important. CONCLUSION: This study showed that dose reduction or withdrawal of palbociclib had no harmful effects in Japanese patients. Efficacy was also high in older patients. It is important to manage palbociclib administration more safely and appropriately. A combination of dose reduction and withdrawal is key to this therapeutic strategy.

A chemoproteoinformatics approach demonstrates that aspirin increases sensitivity to MEK inhibition by directly binding to RPS5.

MEK inhibitors are among the most successful molecularly targeted agents used as cancer therapeutics. However, to treat cancer more efficiently, resistance to MEK inhibitor-induced cell death must be overcome. Although previous genetic approaches based on comprehensive gene expression analysis or RNAi libraries led to the discovery of factors involved in intrinsic resistance to MEK inhibitors, a feasible combined treatment with the MEK inhibitor has not yet been developed. Here, we show that a chemoproteoinformatics approach identifies ligands overcoming the resistance to cell death induced by MEK inhibition as well as the target molecule conferring this resistance. First, we used natural products, perillyl alcohol and sesaminol, which induced cell death in combination with the MEK inhibitor trametinib, as chemical probes, and identified ribosomal protein S5 (RPS5) as their common target protein. Consistently, trametinib induced cell death in RPS5-depleted cancer cells via upregulation of the apoptotic proteins BIM and PUMA. Using molecular docking and molecular dynamics (MD) simulations, we then screened FDA- and EMA-approved drugs for RPS5-binding ligands and found that acetylsalicylic acid (ASA, also known as aspirin) directly bound to RPS5, resulting in upregulation of BIM and PUMA and induction of cell death in combination with trametinib. Our chemoproteoinformatics approach demonstrates that RPS5 confers resistance to MEK inhibitor-induced cell death, and that aspirin could be repurposed to sensitize cells to MEK inhibition by binding to RPS5.

Histopathological findings of an exposed human pulp carbonised by CO2 laser irradiation: A case report.

This report shows the healing process of an exposed pulp carbonised by CO2 laser irradiation prior to the application of a capping material. Six intact teeth from four volunteers were irradiated by CO2 laser and randomly capped with either an adhesive resin (SE bond) (n = 3) or calcium hydroxide-based cement (Dycal) (n = 3). The laser was operated in super-pulsed mode (power output, 0.5 W) for an irradiation time of 30 s. All cavities were restored with composite resin. Each tooth was extracted at approximately 30, 50 or 260 days post treatment and prepared for histological evaluation. CO2 laser irradiation controlled exudate and bleeding from each exposed pulp. Histological images revealed Dycal promoted complete dentine bridge formation at the carbonised pulp surface, and laser energy affected not only the pulp surface but also the deeper part of the pulp chamber.

[Evaluated Cases of Anastomotic Recurrence in Colorectal Cancer].

Anastomotic recurrence in 6 colorectal cancer cases during the postoperative surveillance between 2008 and 2015 was evaluated retrospectively. Five cases had undergone DST anastomosis for sigmoidectomy and proctectomy. They had a pathological tendency to have T3 tumor and deeper, positive lymph node metastases and positive lymphatic and vascular invasion. There were 2 cases with the anastomotic recurrence diagnosis 6 to 8 months after the primary tumor resection while 4 resected cases had recurrent tumor depth of T3, though 3 cases were diagnosed 1 year after the primary tumor resection. Anastomotic recurrence should be considered a few months after primary tumor resection.

[A Case of Laparoscopic Distal Gastrectomy after Abdominal Incisional Hernia Repair].

Re-laparotomy with resection of the mesh after abdominal incisional hernia repair may cause recurrence of the hernia and infection of the mesh. In the present study, we performed laparoscopic distal gastrectomy(LDG)for early gastric cancer without the resection of the mesh in such a case. A 82-year-old man who had undergone abdominal vascular replacement, cholecystectomy, abdominal incisional hernia repair with the mesh, sigmoidectomy had local recurrence of gastric cancer after endoscopic submucosal resection. We diagnosed as cStage IA and performed LDG without resection of the mesh. He had no recurrence of hernia nor infection of the mesh. Minimizing damage to the abdominal wall by laparoscopic surgery can prevent them.

[A Case of a Retroperitoneal Liposarcoma with Long-Term Survival after Four Surgical Resections].

Retroperitoneal liposarcoma is a relatively rare tumor. The only established therapy is surgical resection and the tumor often recurs. This paper deals with a case of a retroperitoneal liposarcoma in which frequent surgical resections for recurrent tumors have provided relatively long-term survival for the patient. The patient was a 70-year-old woman who had undergone surgical resection for a right retroperitoneal tumor. The pathological diagnosis was dedifferentiated liposarcoma. Thereafter she experienced frequent recurrences which required 3 surgical resections. By means of positive margin for the last surgery, chemotherapy with eribulin was administered. There has been no recurrence 13 months after the last surgery.

[A Case of Resection for Metastatic Osseous Tumor from Rectal Cancer].

A 67-year-old male was referred to our hospital for further investigation of fecal occult blood. We diagnosed him with rectal cancer with osseous metastasis. Chemo-and radiation therapy were administered following resection of the rectal cancer. There were no other lesions except for the osseous metastasis remaining after these interventions. The osseous lesion was then resected. There have no signs of recurrence for 1 year and 9 months since the last operation. We report a case of successful resection of osseous metastasis from rectal cancer.

A case of a horseshoe appendix.

Anomalies of the appendix are extremely rare, and a horseshoe appendix is even rarer. A literature search has revealed only five reported cases. In this report, we present a case of a horseshoe appendix.A 78-year-old man was referred for further examination following a positive fecal occult blood test. A mass in his ascending colon was detected on colonoscopy, while computed tomography showed that it was connected to the appendix. Tumor invasion derived from the ascending colon or appendix was suspected. We diagnosed ascending colon cancer prior to laparoscopic ileocecal resection. Macroscopic findings showed that the appendix connected to the back side of the mass, while microscopic findings showed that the mucosa and submucosa were continuous from the appendiceal orifice in the cecum to the other orifice in the ascending colon, where a type 1 tumor was observed on the orifice. We eventually diagnosed the patient with tubulovillous adenoma and a horseshoe appendix.A horseshoe appendix communicates with the colon at both ends and is supplied by a single fan-shaped mesentery. Cases are classified by the disposal of the mesentery and the location of the orifice. Anatomical anomalies should be considered despite the rarity of horseshoe appendices.

Histological evaluation of direct pulp capping of rat pulp with experimentally developed low-viscosity adhesives containing reparative dentin-promoting agents.

OBJECTIVES: This study examines the wound healing process in exposed rat pulp when capped with experimental adhesive resin systems. METHODS: Experimental adhesive resin system for direct pulp capping was composed of primer-I (PI), -II (PII), and -III (PIII) and an experimental bonding agent (EBA). PI was Clearfil(®) SE Bond(®)/Primer (CSP) containing 5.0 wt% CaCl2, PII was PI containing 10 wt% nanofiller (Aerosil(®) 380), and PIII was CSP containing 5.0 wt% of compounds of equal moles of synthetic peptides (pA and pB) derived from dentin matrix protein 1. EBA was Clearfil(®) SE Bond(®)/Bond (CSB) containing 10 wt% hydroxyapatite powders. Three experimental groups were designed. PI was assigned to experimental Groups 1 and 3. PII was assigned to experimental Groups 2 and 3. PIII and EBA were assigned to all experimental adhesive groups. Control teeth were capped with calcium hydroxide preparation (Dycal(®)), and CSP and CSB were applied to the cavity. The rats were sacrificed after each observation period (14, 28, 56, and 112 days). The following parameters were evaluated: pulp tissue disorganization, inflammatory cell infiltration, reparative dentin formation (RDF), and bacterial penetration. RESULTS: There were no significant differences among all the groups for all parameters and all observation periods (p>0.05, Kruskal-Wallis test). All groups showed initial RDF at 14 days postoperatively and extensive RDF until 112 days postoperatively. Groups 2 and 3 demonstrated higher quantity of mineralized dentin bridge formation compared with Group 1. CONCLUSIONS: Addition of nanofillers to the primer was effective in promoting high-density RDF. CLINICAL SIGNIFICANCE: Experimentally developed adhesive resin systems induce the exposed pulp to produce almost the same quantity of reparative dentin as calcium hydroxide. However, we need further studies to elucidate whether the same results could be obtained in humans.

Bond strength of composite resin to enamel and dentin prepared with Er,Cr:YSGG laser.

We aimed to examine the effect of various adhesive systems on the bond strength of composite resin to enamel or dentin prepared with erbium, chromium: yttrium scandium gallium garnet (Er,Cr:YSGG) laser. Each laser-cut enamel or dentin surface was treated with a bonding agent (SBB, Group 1); self-etching primer (SBP) and SBB (Group 2 and control); phosphoric-acid (KET), SBP and SBB (Group 3); KET, sodium-hypochlorite (ADG), SBP and SBB (Group 4); all-in-one adhesive (TSB, Group 5); or KET, ADG and TSB (Group 6). The control group was only polished with wet silicon carbide papers. The enamel shear bond strength of Group 5 was significantly lower than that of other groups (p<0.01). The control group showed higher bond strength compared to Groups 1-6 (p<0.05). Preconditioning using phosphoric acid or phosphoric acid followed by sodium hypochlorite increased the bond strength of composite resin to enamel and dentin prepared using an Er,Cr:YSGG laser.

Conditioning effects of cavities prepared with an Er,Cr:YSGG laser and an air-turbine.

The purpose of this study was to examine, morphologically and histochemically, five types of conditioning effects on cavities prepared with an Er,Cr:YSGG laser and an air-turbine. Cavities were prepared using a Waterlase(®) MD turbo handpiece (W) and an air-turbine (AT) on human extracted molars. The cavity conditionings used were non-conditioned (G1), K-etchant Gel (G2), K-etchant Gel + AD Gel (G3), Clearfil SE Bond primer (G4) and Clearfil S(3) Bond (G5). On naked eye observations, enamel of G1, G2 and G3 in the W cavities and etched enamel of G2 and G3 in the AT cavities were observed as rough and dull in appearance. G4 and G5 in W and AT cavities were observed as shiny surfaces. On SEM observations, no smeared layer was observed in W cavities, while a smeared layer and bur-scratches were observed in AT cavities. In W cavities, rough surfaces were observed on enamel. That is, cracks and minute rough surfaces were observed. In contrast, equally etched scale-shaped enamel rods were observed in AT cavities. Widely opened dentinal tubules and protruding peritubular matrices of dentin were observed in W cavities. A few remaining smeared plugs could be observed at the AT cavities. On LM observations, 13-16 µm layers of the dentin in G1, G2, G4 and G5 of W cavities were stained red in color by the Azan staining method, while redness was not observed in G3. No groups were stained red in AT cavities. It was considered that layers stained red in color were thermal degeneration layers of dentin induced by W. Namely 30 s etching of 40% phosphoric acid gel followed by 90 s treatment of 10% NaClO gel should be recommended for use when combined with an Er,Cr:YSGG laser for cavity preparation.

Histopathological and immunohistochemical study on the effects of a direct pulp capping experimentally developed adhesive resin system containing reparative dentin-promoting agents.

The studies so far conducted by our colleagues in relation to dental pulp capping using adhesive resins revealed that the adhesive resins are useful for capping exposed pulps but are a little slower to take effect on the injured pulp tissue during the initial stage: up to 90-days after pulp exposure, compared with calcium hydroxide and its preparations. In the present study, an experimentally developed adhesive resin system was applied in direct pulp capping and restoration and the healing process was examined histopathologically and immunohistochemically. The resin system was manufactured with calcium phosphate added into the bonding material for the purpose of accelerating the healing process. The largest amount of reparative dentin was formed by SE5 (whitlockite 5 wt%), followed by SE9 (hydroxyapatite 5 wt%, whitlockite 5 wt%), SE1 (hydroxyapatite 5 wt%), and SE2 (hydroxyapatite 10 wt%). Generally, it could be said that the experimental groups using whitlockite and hydroxyapatite had the tendency to produce a larger amount of reparative dentin.

[Analysis of extended-spectrum CTX-M-14 beta-lactamase (ESBLs) producing Escherichia coli isolates in the same ward over the long term].

We bacteriologically and genetically analysed 30 cephalosporin-resistant Escherichia coli strains isolated from specimens from 19 neurology-ward inpatients at our hospital over the 3 years from April 2006 to March 2009, surveying and comparing subjects' backgrounds. Of the 30, 19 (63%) were urine, 6 (20%) sputum, and 3 (10%) blood. We tested extended-spectrum beta-lactamase (ESBLs) production, found in all samples. PCR and gene sequencing showed that 25 strains (83%) were CTX-M-14 and 5 (17%) CTX-M-2. Among CTX-M-14 strains, two cluster groups I and II, were obtained using pulsed-field gel electrophoresis (PFGE). Cluster group I in particular, continued to be detected for 18 months in the same hospital room. The detection rate was high at 13 (68%) in subjects with urinary catheters and morbidity was high in those with a history of cerebrovascular disease, diabetes, and hypertension. Our findings suggest that genetically identical strains may become established and spread in hospitals possibly due to inadequate contact prevention, subjects' immune status, and risk factor existence.

Direct pulp capping effect with experimentally developed adhesive resin systems containing reparative dentin-promoting agents on rat pulp: mixed amounts of additives and their effect on wound healing.

This study examined the wound-healing process of exposed rat pulp when treated with experimental adhesive resin systems. The experimental direct pulp capping adhesive resin systems were composed of primer-I, primer-II and an experimental bonding agent. Primer-I was Clearfil SE Bond (CSE) primer containing 1.0 or 5.0 wt% CaCl(2), and primer-II was CSE primer containing 0.1, 1.0 or 5.0 wt% compound of equal mole of pA and pB with synthetic peptides derived from dentin matrix protein 1 (DMP1). Primer-I containing 1.0 and 5.0 wt% CaCl(2) was assigned to the experimental groups 1-3 and 4-6, respectively. Primer-II containing 0.1, 1.0 or 5.0 wt% compound of pA and pB was assigned to the experimental groups 1 and 4, 2 and 5, and 3 and 6, respectively. In all experimental groups, CSE bond containing 10 wt% hydroxyapatite powder was used as the experimental bonding agent. The positive control teeth were capped with calcium hydroxide preparation (Dycal), and the negative control teeth were capped with CSE. The specimens were alternately stained with Mayer's H&E and the enhanced polymer one-step staining methods. Experimental groups 1, 4, 5 and 6 showed a higher level of reparative dentin formation compared to the negative control 14 days postoperatively. At 28 days postoperatively, all experimental groups showed the formation of extensive reparative dentin, and experimental groups 4, 5 and 6 demonstrated similar dentin bridge formation as that of the positive control. How quickly reparative dentin formation occurs may depend on the concentration of CaCl(2) and the compound of pA and pB in the experimental primer.