Nasa Morokawa

Expression of progastrin-releasing peptide and gastrin-releasing peptide receptor mRNA transcripts in tumor cells of patients with small cell lung cancer.

Abstract Purpose. Small cell lung cancer (SCLC) is a rapidly growing neoplasm accounting for approximately 20% of patients with lung cancer. Progastrin-releasing peptide (proGRP) is produced in about two-thirds of SCLC tumors and is used as a specific marker for SCLC. Although GRP is known to have a variety of biological functions, only limited information is available concerning expression of proGRP mRNA and protein, and that of the receptor for GRP (GRPR) in SCLC tumors. Methods. In individuals with SCLC, the levels of serum proGRP(31–98) were measured by enzyme-linked immunosorbent assay. Expression of proGRP as well as GRPR mRNA in SCLC tumor tissues was investigated by reverse transcription-nested polymerase chain reaction (PCR) amplification. The proportions of alternatively spliced proGRP mRNA transcripts were analyzed in proGRP-producing tumors by nested and competitive PCR amplification. Finally, production of proGRP protein in SCLC tumor was evaluated by using immunohistochemical staining with a polyclonal human anti-proGRP antibody. Results. ProGRP mRNA transcripts could be detected only in tumor tissues recovered from individuals with high serum proGRP levels. The proportions of mRNA subtypes in each case were nearly the same, revealing type I of 55.4±7.6%, type II with 21-b deletion of 1.8±3.6%, and type III with 19-b deletion of 42.8±4.3%, respectively. ProGRP protein production was demonstrated in tumor tissues exclusively from individuals exhibiting high serum proGRP levels. In contrast, GRPR mRNA transcripts were detectable in cancer cells from two of five proGRP-expressing tumor tissues. Conclusions. ProGRP mRNA expression is closely related with the synthesis of proGRP protein which is eventually released into the blood. It is suggested GRP may function as an autocrine growth factor for cancer cells in a subgroup of SCLC patients through, at least in part, upregulation of GRPR expression.

CFTR gene mutations in Japanese individuals with congenital bilateral absence of the vas deferens

Congenital bilateral absence of the vas deferens (CBAVD) is a monosymptomatic disease confined to the male reproductive system with similarity to the phenotype of cystic fibrosis (CF), and mutations in the CFTR gene are highly prevalent in Caucasian CBAVD patients. While CF is very rare in Japan, CBAVD is not. Our previous study demonstrated high prevalence of the 5T allele in the CFTR gene in Japanese CBAVD patients. We analyzed whole exons of the CFTR gene in 19 CBAVD patients and 53 normal individuals using polymerase chain reaction amplification-single strand conformation polymorphism analysis and direct sequencing. Three missense mutations (W216X, G1349S, Q1352H) were found in seven CFTR alleles, and the 5T allele was positive in 11 of 38 CFTR patient alleles. Consequently, 47% of CFTR chromosomes in the patients were affected, and 11 individuals (58%) had at least one mutated CFTR allele. In contrast, three of 53 normal individuals (5.7%) had a missense mutation in one of the CFTR genes, but no 5T allele was detected (both P<0.0001). Mutations of the CFTR gene are closely associated with Japanese patients with CBAVD.

A Japanese patient homozygous for the H1085R mutation in the CFTR gene presents with a severe form of cystic fibrosis

To the Editor: Cystic fibrosis (CF), the most common autosomal recessive disorder in Caucasians, has long been believed to be extremely rare in Orientals (1, 2). In Japan, there have been approximately 130 cases of CF reported in the literature during the last 5 decades, with the estimated incidence of one in 350000 live births (3). Besides, very limited information is available about the spectrum of CF transmembrane conductance regulator (CFTR) gene mutations among Japanese CF patients, although more than 800 mutations of the CFTR gene, including the most common DF508, have been accumulated worldwide through the CF Genetic Analysis Consortium [(4), personal communication]. Here we describe a 15-year-old female Japanese patient who exhibited typical manifestations of CF and proved to be a homozygote for a very rare missense mutation H1085R. She was born to consanguineous parents at full term. From 1 year old, she repeatedly suffered from respiratory symptoms associated with Pseudomonas aeruginosa infection and experienced occasional steatorrhea. At 8 years old, she developed fecal ileus and underwent surgical operation. At 9 years old, she was diagnosed as CF based on the elevated sweat chloride level at 201 mmol/l. She also had chronic sinusitis, and abdominal ultrasonography revealed fibrotic changes of the pancreas. The patient has recently developed shortness of breath on exertion, and room-air arterial blood gas analysis showed moderate hypoxemia. For evaluation of the CFTR gene mutations, her DNA sample was evaluated for 32 major CFTR gene mutations responsible for more than 90% of CF chromosomes in the Caucasian population of North America (5). However, none of these mutations were detected in her CFTR alleles. Next, all 27 exons of the CFTR gene, including both the 5%and 3%-intron–exon junctions, were polymerase chain reaction (PCR) amplified and studied by single strand DNA conformation polymorphism (SSCP) analysis (6, 7). When the PCR products were electrophoresed and silver-stained, the DNA bands with different mobility from those of the control were only detected in exon 17b. Direct sequencing of the band clearly demonstrated an alteration of the nucleotide residue at 3386 from A to G in a homozygous fashion, resulting in His to Arg change at the amino acid 1085, a missense mutation H1085R. The CFTR genotype analysis of her family by PCR amplification of exon 17b and subsequent RsaI restriction digestion revealed that both parents and her three siblings were heterozygous for the mutation (Fig. 1). Further evaluation for three polymorphic loci proved that the H1085R mutation allele was associated with a haplotype of (GATT)6 in intron 6a (8), (TG)12T7 in the TG repeat and polythymidine tract at the splice acceptor site in intron 8 (9, 10), and M470 (1540A) in exon 10 (11, 12), respectively. The mutation H1085R, first described by Mercier et al. (13, 14), was detected on one of the CFTR alleles of a French CF patient who was pancreatic insufficient and a compound heterozygote with DF508 on the other chromosome [(13), personal communication]. However, no other CF patients with H1085R have been reported since then, implying that it is an extremely rare mutation (14). Consistent with the previous report that other mutations located in exon 17b, such as R1066L and M1101R, were usually associated with pancreatic insufficiency, the case presented here and the French case had pancreatic insufficiency, suggesting that the H1085R is also a severe allele [(13), personal communication]. Since the amino acid residue 1085 is located between the fourth and the fifth transmembrane segments of the second membrane-spanning domain and resides at the gate of the channel in the cytoplasm, it is conceivable that a His to Arg alteration would profoundly affect Cl conductance by either reduced Cl flow through a single open channel or by shortened duration for the channel opening (1, 15). One important point is that the H1085R alleles of these 2 patients are most likely to be recurrent

Marked Improvement with Pirfenidone in a Patient with Idiopathic Pulmonary Fibrosis

A man in his mid-60s with idiopathic pulmonary fibrosis and hepatitis B-related liver cirrhosis developed exertional dyspnea and a dry cough lasting for three months. High-resolution computed tomography (HRCT) showed increasing bilateral ground-glass opacity superimposed on the usual interstitial pneumonia pattern. Six months after starting pirfenidone therapy, the partial pressure of arterial oxygen at rest increased from 81 to 101 torr, the predicted forced vital capacity (FVC) value increased from 75% to 94% and the ground-glass opacity on HRCT improved. The FVC value was subsequently maintained near or above baseline for 43 months. We concluded that our patient was a super-responder to pirfenidone therapy.

The Efficacy and Safety of Long-term Pirfenidone Therapy in Patients with Idiopathic Pulmonary Fibrosis

Objective Pirfenidone (PFD) is often used for years, but the efficacy and safety of long-term PFD therapy in patients with idiopathic pulmonary fibrosis (IPF) are not fully understood. Methods and Patients We retrospectively evaluated 46 patients with IPF who received PFD between February 2009 and August 2014. The efficacy and safety of PFD therapy were compared between 2 groups: long-term therapy patients who received PFD for over 1 year (group L, n=30, 65%) and short-term therapy patients who could not receive PFD for more than 1 year due to worsening of their condition or side effects (group S, n=16, 35%). Results The median age of the 46 patients was 70.5 years, and the median baseline % predicted forced vital capacity (%FVC) was 70.0%. The changes in the FVC in group L were -120 mL and -170 mL at 12 and 24 months after receiving PFD, respectively. The respective median survival times after PFD therapy in groups L and S were 1,612 days and 285 days (p<0.001). The patients in group L experienced a longer time free of acute exacerbation of IPF than those in group S (947 days vs. 145 days, p=0.001). A multivariate analysis revealed that %FVC <60% was a predictor of the inability to receive PFD for over 1 year (odds ratio 0.240, 95% confidence interval 0.060-0.958; p=0.043). With regard to grade 3-5 adverse events, only one patient exhibited grade 3 hyponatremia. Conclusion Long-term PFD therapy is effective, with few severe adverse events.

Antemortem diagnosis of pulmonary tumor thrombotic microangiopathy in a patient with recurrent breast cancer: a case report

BackgroundPulmonary tumor thrombotic microangiopathy (PTTM), a rare complication of advanced cancer, is histologically characterized by tumor embolisms and fibrocellular intimal proliferation of small pulmonary arteries and arterioles. PTTM usually has an extremely poor prognosis, and antemortem diagnosis is very difficult.Case presentationA 65-year-old woman with a 5-year history of clinical stage IIA (T2N0M0) invasive ductal carcinoma of the left breast was hospitalized for worsening shortness of breath, hemoptysis, and cough since 2 months. She had previously received neoadjuvant chemotherapy and left mastectomy. Because the cancer cells were positive for human epidermal growth factor receptor 2 (HER2), four cycles of trastuzumab had been administered as adjuvant chemotherapy. On admission, chest computed tomography (CT) showed peripheral consolidations in both the lower lobes and a mediastinal mass. Specimens obtained on video-assisted thoracoscopic surgical biopsy revealed tumor cell embolism, intimal fibrocellular proliferation of small arteries, fibrin thrombi, recanalization, and infarction in the left lower lobe, as well as metastasis to the mediastinal pleura. Immunohistochemical staining of the tumor cells revealed positivity for HER2, and a diagnosis of recurrent breast cancer with PTTM was made. Four cycles of trastuzumab resulted in rapid improvement of her symptoms and CT findings of peripheral consolidations and the mediastinal mass.ConclusionAn antemortem diagnosis of PTTM was made in a patient with HER2-positive recurrent breast cancer. Trastuzumab was effective for not only breast cancer but also PTTM.

Nebulized liposomal amphotericin B for treating Aspergillus empyema with bronchopleural fistula.

Aspergillus empyema is an uncommon condition associated with a high mortality rate (1). Treatment options for this condition include surgical drainage and antifungal therapy, but intravenous antifungal monotherapy is usually inefficient in the advanced stages of the disease, as discussed in several case reports (2–7). Treatment can fail because of insufficient drug delivery and penetration into the pleural space by intravenous administration of antifungal agents. We describe a case of Aspergillus empyema with a bronchopleural fistula and the clinical use of liposomal amphotericin B delivered directly to the infection site using a nebulizer.

OK-432 pleurodesis for the treatment of pneumothorax in patients with interstitial pneumonia

BACKGROUND Pneumothorax occasionally develops in patients with interstitial pneumonia (IP) and is often intractable. As there exists no well-established treatment for pneumothorax with IP, we evaluated the efficacy and safety of pleurodesis with OK-432, a lyophilized preparation of Streptococcus pyogenes Su strain that has been inactivated by benzylpenicillin. METHODS We retrospectively evaluated the efficacy and safety of pleurodesis using OK-432 in 39 patients treated for IP-related pneumothorax between January 2006 and May 2017. Five to 10 Klinische Einheit (KE) of OK-432 was injected through the chest tube of each patient. Pleurodesis was considered successful if 1) the chest tube was removed without air leaks and 2) there was no recurrence of pneumothorax within 4 weeks after tube removal, and no additional treatment was required. RESULTS OK-432 pleurodesis was performed 46 times in 39 patients. The median number of OK-432 intrapleural injections received was 1 (range, 1-6), and median dose was 10 KE (range, 5-55 KE). The success rate was 63% (29/46) and recurrence rate was 17.4% (8/46). Grade 5 adverse events were observed in eight patients, including two patients who developed acute exacerbation of IP. Patients in whom the first OK-432 pleurodesis was successful had a significantly longer median survival time than patients in whom it was unsuccessful (322 days vs. 70 days, p = 0.036). CONCLUSIONS Our results show that OK-432 pleurodesis is an effective treatment for pneumothorax associated with IP; however, clinicians should be aware of the possibility of adverse events, especially in patients who are critically ill.

Cavitary pulmonary infection with Mycobacterium avium observed by bronchoscopy.

A 58-year-old man was admitted to our hospital because of fever and loss of appetite. He had undergone surgery for esophageal cancer. A chest radiography 12 years after the surgery revealed cavitary lesions in the right upper lobe of the lung. The patient was then diagnosed as having Mycobacterium avium infection. The cavitary lesions worsened 2 years after clarithromycin monotherapy. Bronchoscopy was performed to observe the interior of the cavity. Gray debris adhering to the cavitary wall decreased after intensive treatment with Streptomycin, rifabutin, levofloxacin, and ethambutol. This is a rare case in which treatment efficacy of M. avium infection was directly observed by serial bronchoscopy.