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A 75‐year‐old woman underwent thoracoscopic right upper lobectomy for lung cancer. A histopathological examination showed adenocarcinoma, pT1aN0M0 stage IA1. At six months after surgery, chest computed tomography (CT) revealed pericardial nodules that had not been detected before pulmonary resection. Postoperative CT performed two months later revealed that the nodules were growing and F18 fluorodeoxyglucose‐positron emission tomography showed a maximum standardized uptake of 9.87. Blood tests revealed no elevated tumor markers, with the exception of a mildly elevated interleukin‐2. Based on the above results, thoracoscopic biopsy was performed due to the suspected recurrence of lung cancer or malignant lymphoma. The histopathological examination of the nodule revealed immunoglobulin G4 (IgG4)‐related inflammatory pseudotumor. The serum IgG4 levels were elevated (358 mg/dL, normal: 4.5–117.0 mg/dL). No additional treatment was required because all nodules were observed to have disappeared naturally on a follow‐up CT scan performed two months after the surgical biopsy. The patient has been followed‐up for two years without recurrence. Key points Significant findings of the study We report a case of pericardial immunoglobulin G4‐related inflammatory pseudotumor that appeared after right upper lobectomy for lung cancer, and which naturally disappeared without any treatment. What this study adds There was an immunoglobulin G4‐related inflammatory pseudotumor which appeared as multiple nodules in the pericardial space, and this should be kept in mind when considering the differential diagnosis of intrapericardial nodules., We report a case of pericardial immunoglobulin G4‐related inflammatory pseudotumor that appeared after right upper lobectomy for lung cancer, and which naturally disappeared without any treatment.

The nymphalid butterfly Araschnia burejana and the papilionid butterfly Papilio xuthus exhibit seasonal diphenism comprising spring-morphs that develop from diapause pupae and summer-morphs that develop from non-diapause pupae. The development of seasonal morphs in A. burejana is regulated by the timing of secretion of ecdysteroids for adult development, whereas that in P. xuthus is regulated by the secretion of summer-morph-producing hormone, which is present in the brains and is under control of the photoperiod. We investigated whether a cerebral factor derived from brains plays a significant role in the regulation of seasonal morph development in A. burejana using surgical operations. Pairs of chilled diapause pupae that had been chilled for more than 3 months at 4°C were joined surgically to each other and then developed into spring-morph or spring-like-morph adults. Chilled diapause pupae that were joined with 1-day-old non-diapause pupae developed into summer-morph or summer-like-morph adults. When the brains of non-diapause pupae were removed surgically 6-8 hr after pupation with and without injection of 20-hydroxyecdysone, a large portion of them developed into spring-morph or spring-like-morph adults, respectively. Furthermore, 90% of non-diapause pupae developed into spring-morph or spring-like-morph adults when the neck was ligated within 5 min after pupation. These results indicated that a cerebral factor showing summer-morph-producing hormone activity, which is secreted from the brain in the early pupal stage, in addition to 20-hydroxyecdysone for adult development, play a significant role in the determination of summer-morph development in non-diapause pupae of A. burejana.

Field studies of butterflies were carried out in four different selected areas i.e. Krishnopur, AERE campus, Rajalak Farm and Horters (Part of JU campus) at Savar, Dhaka. Butterflies identified of those areas constituted 158 species belonging 10 families, 87 species of which are newly-recorded in Bangladesh. Numbers of species identified in each family reached 47 in Lycaenidae (30 newly recorded), 32 in Hesperiidae (22 new), 22 in Pieridae (11 new), 19 in Nymphalidae (6 new), 14 in Papilionidae (7 new), 14 in Saytridae (8 new), 5 in Danaidae (1 new), 3 in Riodinidae (1 new), 1 in Acraeidae and 1 newly recorded Amathusiidae. The maximum 130 species and the minimum 65 species were recorded in Krishnopur and Rajalak Farm, respectively. Almost all butterfly families were found to be dominant in Krishnopur. Among the total identified species, 35.44% belonged to very rare (VR) and 20.25% to rare (R) categories, whereas 12.18%, 12.03% and 13.29% species were very common (VC), common (C) and not rare (NR) categories, respectively. It may be that the numbers of butterfly species as well as their fauna vary greatly depending on the floral and ecological conditions in and around the study areas.DOI: http://dx.doi.org/10.3329/ujzru.v30i0.10713Univ. j. zool. Rajshahi Univ. Vol. 30, 2011 pp. 09-15

Pupae of the painted lady butterfly Vanessa cardui exhibit pupal color polyphenism consisting of white, dark and intermediate types. We investigated environmental factors affecting pupal coloration and the physiological mechanisms underlying the control of pupal color polyphenism in this species. Over 80% of larvae reared at 16 degrees C developed into pupae of dark types, whereas over 82% of larvae at 32 degrees C developed into pupae of white types irrespective of long/short-day photoperiod conditions. When mature larvae reared at 32 degrees C were ligatured between thoracic and abdominal parts at three different pharate pupal stages, all of the head-thoracic parts developed into white pupae regardless of pupal stage, but all abdominal parts ligatured at the early pharate pupal stage only developed into dark pupae. These results indicate that temperature during larval stages is an important element affecting pupal coloration as an environmental cue in V. cardui, and that a factor(s) inducing white pupae is released from head-thoracic parts under conditions of high temperature. Additionally, when ligatured abdomens destined to develop into dark pupae were treated with crude extracts prepared from the central nervous system, all of the ligatured abdomens developed into white pupae at a level dependent on dose and pupal stage. These results suggest that the factor inducing white pupae is a key molecule controlling pupal color polyphenism in V. cardui.

Three species of nymphalid butterflies, Vanessa cardui, V. indica and Nymphalis xanthomelas japonica, do not exhibit seasonal polyphenism in wing coloration. To determine whether seasonal non-polyphenic butterflies possess a cerebral factor affecting wing coloration, we used a Polygonia c-aureum female short-day pupal assay for detection of summer-morph-producing hormone (SMPH) activity in P. c-aureum. When 2% NaCl extracts of 25 brain-equivalents prepared from the pupal brains of V. cardui, V. indica or N. xanthomelas japonica were injected into Polygonia female short-day pupae, all recipients developed into summer-morph adults with dark-yellow wings, and the average grade score (AGS) of summer morphs showing SMPH activity was 3.8, 3.7 and 4.0, respectively. In contrast, when acetone or 80% ethanol extracts prepared from pupal brains were injected into Polygonia pupae, all recipients developed into autumn-morph adults with a dark-brown coloration and each exhibited an AGS of less than 0.5. Our results indicate that a cerebral factor showing SMPH activity is present in the pupal brain of seasonal non-polyphenic nymphalid butterflies, suggesting that a SMPH and cerebral factor showing SMPH activity occur widely among butterfly species. This finding will improve our understanding of the presence of cerebral factors showing interspecific actions of SHPH.

Background Mutations of the epidermal growth factor receptor (EGFR) gene at kinase domain have been reported in non-small-cell lung cancer (NSCLC), and some common somatic mutations in EGFR have been examined for their ability to predict sensitivity to gefitinib or erlotinib. However, EGFR mutations at exon 20 have been reported to predict resistance to gefitinib therapy. Materials and methods We investigated the EGFR mutations and/or polymorphism statuses at kinase domain in 303 surgically treated non-small cell lung cancer (NSCLC) cases. One hundred ninety-four adenocarcinoma cases were included. The presence or absence of EGFR polymorphism of kinase domains was analyzed by direct sequences. We have also investigated EGFR polymorphism status at exon 20 for 23 NSCLC patients who had undergone surgery followed by treatment with gefitinib at the National Hospital Organization, Kinki-chuo Chest Medical Center. Results EGFR mutations at kinase domain were found in 75 of 303 lung cancer patients. During sequencing of EGFR tyrosine kinase domain in tumors, 86 EGFR polymorphism (G2607A) cases were identified at exon 20. G2067A polymorphism was significantly higher in nonadenocarcinomas (37.4%) than in adenocarcinoma (25.3%, P = 0.0415). The polymorphism status did not correlate with gender, smoking (never smoker versus smoker), and EGFR mutations. In 46 total gefitinib treated NSCLC patients, there was a tendency toward better prognosis in EGFR wild type (GG) patients than AG + AA patients. EGFR polymorphism in Japanese lung cancers seemed to be less frequent than Caucasian lung cancers. Conclusions EGFR-tyrosine kinase polymorphism might be associated with clinicopathological background of lung cancers.

Mutations of the epidermal growth factor receptor (EGFR) gene at kinase domain have been reported in non-small-cell lung cancer (NSCLC). However, EGFR mutations status at C-terminal domain has not been reported in detail.We investigated the EGFR mutation and polymorphism statuses at C-terminal domain in 398 surgically treated NSCLC cases. Two hundred and sixty-eight adenocarcinoma cases were included. The presence or absence of EGFR mutation and polymorphism was analyzed by direct sequences.A novel EGFR somatic mutation at exon 25 (G3034, D1012H) was found from 1 of 398 lung cancer patients. During sequencing of EGFR C-terminal domain in NSCLC, 194 EGFR polymorphism (C2982T) cases were identified at exon 25. The polymorphism statuses were not correlated with gender, smoking status (never smoker vs. smoker), pathological subtypes and EGFR mutations. The EGFR polymorphism ratio was significantly higher in younger NSCLC (or =60, 56.8%) than in older NSCLC (60, 45.6%, P = 0.0467). The EGFR polymorphism ratio was significantly higher in lymph node positive NSCLC (57.4%) than in lymph node negative NSCLC (44%, P = 0.0168). In 46 total gefitinib treated NSCLC patients, exon 25 polymorphism was not correlated with prognosis.EGFR mutation at C-terminal in lung cancers seemed to be extremely rare, however, this D1012H mutation might be a role in EGFR function. EGFR polymorphism at exon 25 might be correlated with progression of NSCLC.

ErbB3 is a member of the epidermal growth factor receptor (EGFR) family of tyrosine kinase. A previous report has suggested that overexpression of ErbB3 protein was associated with lung cancer recurrence. Other reports suggested that high ErbB3 protein expression was associated with shorter survival in advanced non-small-cell lung carcinomas (NSCLC). The relationship between the erbB3 mRNA expression and EGFR mutation has not been reported. In this study, we have investigated the erbB3 mRNA levels in reference to clinicopathologic data and EGFR mutation status in Japanese lung cancer specimens.This study included 192 surgically removed NSCLC cases operated at Nagoya City University Hospital. The erbB3 mRNA levels were quantified by real-time polymerase chain reaction using LightCycler (Roche Molecular Biochemicals, Mannheim, Germany).The erbB3/glyceraldehydes-3-phosphatate dehydrogenase levels were significantly higher in female than in male (P0.0001), higher in nonsmoker than in smoker (P = 0.0007), higher in adenocarcinoma than in other types of lung cancers (P0.0001), and higher in well-differentiated than in less-differentiated subtypes (P = 0.0304). The erbB3/glyceraldehydes-3-phosphatate dehydrogenase levels were significantly higher in patients with EGFR gene mutation than in patients without mutation (P = 0.0124).The present study suggests that increased expression of erbB3 may play an additional role in NSCLC especially in female, nonsmoker, adenocarcinoma, and with EGFR gene mutation. Further studies on large sample sizes would help to determine if erbB3 expression is a clinically useful tool to further define patients who will benefit from EGFR TKIs.

Autoimmune regulator (AIRE) is a transcription factor expressed in the thymic medullary epithelial cells (MECs). MECs have been suggested to contribute to tolerance induction by presenting tissue specific antigens to T cells that react to these antigens. AIRE expression in cells other than MECs has not been studied extensively. Here we report four-color flow cytometric and quantitative real-time PCR studies on the expression of AIRE in the lymphoid cells in the thymus and periphery. In the thymus AIRE was expressed in B cells and CD4+CD8+ double positive cells at a low level. In the peripheral blood AIRE was expressed in B cells but not in T cells, and also expressed in CD14+ dendritic cells (DCs)/macrophages and granulocytes at higher levels. AIRE mRNA was expressed higher in B cells than in T cells. Further study is warranted to define the functional role of the AIRE expressed in the lymphoid cells.

To evaluate the epidermal growth factor receptor (EGFR) protein expression, gene mutations and amplification as predictors of clinical outcome in patients with non-small-cell lung cancer (NSCLC) receiving gefitinib, we have performed fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC). We investigated the EGFR amplification and EGFR protein expression statuses in 27 surgically treated non-small-cell lung cancer (NSCLC) cases. These patients experienced relapse after surgery and received gefitinib 250 mg/day. The presence or absence of EGFR mutations of kinase domains was analyzed by genotyping analysis and sequences, and already reported. EGFR mutations were found from 15/27 lung cancer patients. EGFR mutation status was significantly correlated with better prognosis (log-rank test P=0.0023). Smoking status (never smoker vs. smoker, P=0.0032), and pathological subtypes (adenocarcinoma vs. non-adenocarcinoma, P=0.0011), but not EGFR amplification (P=0.1278), were correlated with survival of lung cancers. EGFR IHC results were correlated with FISH results (P=0.0125), but not correlated with prognosis (P=0.7921). Thus, the EGFR gene amplification or protein expression is not a predictor of gefitinib efficacy in Japanese patients with NSCLC. We have also evaluated the EGFR mutation status and clinico-pathological features for 27 NSCLC patients who had undergone surgery followed by treatment with gefitinib at the National Hospital Organization, Kinki-chuo Chest Medical Center. The EGFR mutation status, especially exon19 mutation was correlated with good response to gefitinib than exon 21 point mutation.

Much evidence has accumulated that the epidermal growth factor receptor (EGFR) and its family members are strongly implicated in the development and progression of lung cancers. Recently, erbB4 kinase domain mutations were reported in Korean patients with lung cancer. We hypothesized that erbB4 mutations correlate with clinicopathologic features of lung cancers.The presence or absence of erbB4 kinase domain mutations was analyzed by reverse-transcription polymerase chain reaction amplification and direct sequencing in 105 surgically treated non-small-cell lung cancer cases from Nagoya City University Hospital. Sixty-three adenocarcinoma cases were included. The EGFR mutation status for these 105 samples were already reported. We have investigated erbB4 expression status by immunohistochemistry in 40 non-small cell lung cancer cases.ErbB4 mutation was not found in 105 patients with lung cancer. The EGFR mutation status was significantly correlated with sex (women, 74.2% vs. men, 9.5%; P0.0001), smoking status (never-smokers, 68.8% vs. smokers, 11%; P0.0001), pathologic subtype (adenocarcinoma, 44.4% vs. non-adenocarcinoma, 4.8%; P0.0001), and differentiation status of the lung cancer (well-differentiated, 47.4% vs. others, 14.8%; P = 0.0004). We detected ErbB4 protein positivity in 20 samples. The ErbB4 protein status was not significantly correlated with sex (women, 28.6% vs. men, 54.5%; P = 0.4075), smoking status (never-smokers, 69.4% vs. smokers, 16.9%; P0.0001), pathologic subtype (adenocarcinoma, 46.2% vs. nonadenocarcinoma, 51.9%; P0.9999), or differentiation status of the lung cancer (well-differentiated, 54.5% vs. others, 48.3%; P0.9999).Thus, erbB4 mutations are rare in Japanese people with lung cancer and of limited value for molecular-targeted therapy.

Purpose Human MOB1 ( hMOB1 ) is a recently isolated gene that is a human homologue of the Schizosaccharomyces mitotic checkpoint gene MOB1 . The loss of checkpoint control in mammalian cells results in genomic instability, leading to the amplification, rearrangement, or loss of chromosomes, events associated with tumor progression. We hypothesized that hMOB1 might be expressed in non–small-cell lung cancer (NSCLC). Patients and Methods We attempted to determine the influence of hMOB1 expression on clinicopathologic features in patients with NSCLC who had undergone surgery. Expression of hMOB1 messenger RNA (mRNA) was evaluated by reverse transcription-polymerase chain reaction in 60 NSCLCs and adjacent histologic normal lung samples using LightCycler®. Results Human MOB1/glyseraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA expression was significantly decreased in the tumor of lung cancer tissue (3.347 ± 4.306) compared with normal lung tissue (4.833 ± 4.306; P = 0.0437), although 22 of 60 lung cancer tissue samples had > 1 tumor-normal ratio of MOB1/GAPDH mRNA expression. There was no relationship between hMOB1 gene expression and age, sex, pathologic stages, or pN status. However, decreased hMOB1/GAPDH expression was especially seen in pT1 lung cancer (tumor-normal ratio; 0.318 ± 0.328) when compared with pT4 lung cancer (1.915 ± 1.895; P = 0.0362). Conclusion The decreased expression of hMOB1 mRNA might be the early phase phenomenon for tumor invasion from NSCLC. Alternatively, loss of mitotic checkpoint might play a role in oncogenesis for lung cancer.

ObjectiveThe presence of epidermal growth factor receptor (EGFR) mutations has been reported to predict the response to gefitinib in pulmonary adenocarcinoma patients. A retrospective analysis was conducted to identify the correlation between computed tomographic findings of the nodules and EGFR status.Patients and MethodsComputed tomographic findings of 38 patients with peripheral pulmonary adenocarcinoma with EGFR mutations were reviewed and compared with those of 42 peripheral pulmonary adenocarcinoma patients with wild-type EGFR.ResultsMutations were found significantly more frequently among women (28 of 45 women versus 10 of 35 men) and among non-smokers (31 of 47 non-smokers and 7 of 33 smokers). The L858R mutation was found in 18 cases. Several types of deletion mutants in exon 19 were found in 18 cases. The nodules with EGFR mutations (2.5 ± 1.0 cm) were significantly smaller in diameter than those in the wild-type group (3.1 ± 1.9 cm). Ground glass opacity (GGO) was more often observed in the mutation group (28 of 38) than in the wild-type group (24 of 42), but the difference was not statistically significant. When mutations were analyzed with reference to both the tumor size and GGO ratio, patients with a tumor ≤3 cm and a GGO ratio ≥50% often had EGFR mutations, and most (10 of 12) were expressed in female patients. No male adenocarcinoma patients with a tumor larger than 4.0 cm had EGFR mutations.ConclusionsEGFR mutations were found most frequently in small peripheral adenocarcinomas with a GGO ratio ≥50%, especially among women. These factors may be useful in deciding therapeutic strategies for adenocarcinomas when resection or biopsy is not feasible.

Background: Epidermal growth factor receptor (EGFR) gene mutations have been reported to correlate with the sensitivity to the tyrosine kinase inhibitor treatment for advanced lung cancers. Since several reports have shown that invasive thymoma overexpress the EGFR protein, we examinedtheEGFRexpressionandmutationstatusesinthymomaandthymiccarcinomatissues. Methods: EGFR mutation statuses from 99 thymic epithelial tumor samples were evaluated by a rapid and sensitive TaqMan assay using Applied Biosysytems 7500 real-time PCR system. Probes were designed according to the 13 different EGFR mutations reported previously in lung cancers. A total of 38 thymoma samples were directly sequenced for the EGFR gene. Protein expressions were evaluated for 56 thymic epithelial tumors by immunohistochemistry. Results: EGFR gene mutations were not detected in any of the thymoma and thymic cancer samples using TaqMan PCR assay. Of the 38 samples 3 showed a heterozygous silent mutation without changes in the protein, a G to A transition at the nucleotide 2361 in exon 18. EGFR expression was significantly higher in invasive thymomas (stages III–IV, 15/19 were positive) than in early stage thymomas (stages I–II, 7/33 were positive) (P < 0.0001). All four carcinomas and all seven B3 thymomas showed EGFR positive staining. Conclusions: Although EGFR mutation at the tyrosine kinase domain is unlikely to be a therapeutictargetforthymoma,theinformationaboutEGFRexpressionwouldcontributetothefurther identification of the therapeutic target for advanced thymomas.

Diapause pupae of the swallowtail butterfly Papilio xuthus L. exhibit diapause-green, orange and brownish-orange color polymorphism. Development of orange pupae involves a neuroendocrine factor inducing orange pupa (Orange-Pupa-Inducing Factor, OPIF), which is secreted from the head–thoracic region during late pharate pupal stages, in particular from the ganglia of short-day animals located posteriorly from the second thoracic ganglion2 (TG2). This report describes certain properties of OPIF using bioassays involving ligated abdomens of short-day pharate pupae. Localization of OPIF in the central nervous system of short-day larvae indicated that it was present predominantly in TG2, thoracic ganglion3 (TG3) and abdominal ganglion1 (AG1) complexes. OPIF activity in TG2,3–AG1 complexes was over two times higher than in the more posteriorely located ganglia. The developmental profile of OPIF in last instar short-day larvae revealed that OPIF activity in larval ganglia posterior to TG2 became gradually higher as larval growth proceeded, suggesting that OPIF might be accumulated in TG2,3–AG1–7 complexes as larvae prepare for pupal molting. Furthermore, ligated abdomens of short-day larvae developed into pupae of an orange type when a 2% NaCl extract containing OPIF prepared from TG2,3–AG1–7 complexes of long-day larvae was injected into ligated abdomens of short-day pharate pupae, indicating that OPIF is also present in long-day larvae. Additionally, a biochemical investigation using gel filtration chromatography showed that the molecular weight of OPIF was about 10 kDa.

Bombyx mori chitinase (Bm-CHI), with a molecular mass of 75 kDa, was investigated on the possibility that it can serve as a biocontrol agent against the adult Japanese pine sawyer (JPS), Monochamus alternatus (Coleoptera: Cerambycidae). Oral ingestion of purified chitinase at concentrations of 3 microM (11.25 microg/50 microl) and 0.3 micoM (1.125 microg/50 microl) caused high mortality in JPS, a significant decrease in bark consumption, and, only in high concentration, a slight reduction of body weight. Fluorescence assays indicated that peritrophic membrane (PM) chitin is degraded by the action of orally ingested Bm-CHI at 3 microM concentration only. Scanning electron micrographs clearly indicated that the beetles that ingested Bm-CHI of the same high concentration had their PM perforated and disrupted, but ultrastructural studies showed that the ingested chitinase did not affect the midgut epithelium. These findings open up the possibility of using insect chitinase as a biopesticidal enzyme. It should have agronomic potential for insect control.

The cabbage armyworm, Mamestra brassicae, has winter-and aestival- diapause pupae (WD- and AD-pupae) showing differences in the strength of diapause. We tried to quantify diapause-strength by measuring the doses of 20-hydroxyecdysone (20-E) required to induce adult development in WD-, AD- and decerebrated non-diapause pupae (ND-pupae). The role of the brain in the regulation of diapause-strength was studied through the decerebration and brain-reimplantation of WD-and AD-pupae. The 20-E doses required for adult development were small within the first 2 days of pupation, and increased thereafter to reach a constant level about 10 days after pupation in AD- and decerebrated ND-pupae. The required 20-E doses in WD-pupae increased for more than 40 days after pupation. When 0-day-old WD- and 0-day-old AD-pupae were decerebrated, required 20-E doses increased after pupation and reached a constant about 10 days later. The required 20-E dose reached a constant level in decerebrated WD-pupae that was smaller than that observed for decerebrated ND- and WD-pupae. Furthermore, the required doses increased when 0-day-old WD-pupal brains were reimplanted into decerebrated WD- and decerebrated ND-pupae. In WD-, AD- and decerebrated ND-pupae, diapause-strength can be represented as the 20-E dose required for adult development. Diapause-strength is weak after pupation, increases thereafter, and reaches a constant about 10 days later in AD- and decerebrated ND-pupae. In WD-pupae, diapause-strength increases for more than 40 days after pupation and reaches a level that is twice that estimated for AD-pupae. Brains of diapausing WD-pupae may secrete a factor that suppresses the 20-E responsiveness of pupal organs, for the purpose of maintaining winter-diapause.

Much evidence has accumulated that the epidermal growth factor receptor (EGFR) and its family members are strongly implicated in the development and progression of lung cancers. Somatic mutations of the EGFR gene were found in about 25–40% of Japanese lung cancer patients. More recently, erbB2 mutations are found in about 4% of European-derived lung cancer patients. We have investigated EGFR and erbB2 mutation status in 95 surgically treated nonsmall cell lung cancer (NSCLC) cases from Nagoya City University Hospital. Seventy-five adenocarcinoma cases were included. The presence or absence of EGFR and ernB2 mutations of kinase domains were analyzed by reverse transcription polymerase chain reaction (RT-PCR) amplifications and direct sequences. We have also investigated erbB2 mutation status in 27 surgically treated NSCLC cases followed by treatment with gefitinib from Kinki-chuo Chest Medical Center. EGFR mutations (CTGfiCGG; L858R) were found from 14 of 95 lung cancer patients. We also detected the deletion 1a-type mutations from 9 patients and deletion 4-type mutations from 6 patients in exon 19. In exon 20, 4 mutations including 2 novel mutations were found. Total EGFR mutations were present in 35 patients (36.8%). These mutation statuses were significantly correlated with gender (women 73.3% vs. men 20%, p < 0.0001), smoking status (never smoker 69.4% vs. smoker 16.9%, p < 0.0001), pathologic subtypes (adenocarcinoma 45.1% vs. nonadenocarcinoma 12.5%, p 5 0.0089) and differentiation status of the lung cancers (well 51% vs. moderately or poorly 18.4%, p 5 0.0021). On the other hand, erbB2 mutation was only found from 1 of 95 patients, at exon 20. This patient was female and a never smoker with adenocarcinoma. This 12 nucleotide insertion mutation (2324–2325 ins ATACGTGATGGC) was located in the exon 20 at kinase domain (775–776 ins YVMA). There was no erbB2 mutation in 27 gefitinib-treated NSCLC patients. In total, we have found only 1 erbB2 mutation from 122 (0.8%) Japanese NSCLC patients. There was a significantly higher erbB2 positive (21/31) ratio in EGFR mutant patients (13/25, 52.0%) compared to EGFR wild-type patients (10/ 62, 16.1%; p 5 0.0247). The NSCLC specimen with erbB2 mutation showed 11 immunoreactivity. The EGFR mutation status might correlate with the clinicopathologic features related to good response to gefitinib, such as gender, smoking history and pathologic subtypes of lung cancers. However, erbB2 mutation is rare from Japanese lung cancer and is of limited value for molecular target therapy. ' 2005 Wiley-Liss, Inc.

Diapause pupae of Papilio xuthus show color polymorphism, represented by diapause-green, orange, and brownish-orange types that are each associated with specific pupation sites. We investigated the role of the site of pupation on the induction of the development of orange types (or brownish-orange types), and the endocrine mechanism underlying the control of color polymorphism in short-day pupae. All short-day larvae of the wandering stage developed into orange or brownish-orange type pupae when they were placed in rough-surfaced containers after gut-purge. Utilizing a pharate pupal ligation between the thorax and abdomen, the endocrine mechanism underlying the control of color polymorphism was shown to involve a head-thorax factor (Orange-Pupa-Inducing Factor: OPIF) that induced orange types in short-day pupae. OPIF was bioassayed using the ligated abdomens of short-day pharate pupae. OPIF was extractable with 2% NaCl solution from 5th-instar larval ganglia complexes following the mesothoracic complex (TG(2,3)-AG(1-7)), but it could not be extracted with either acetone or 80% ethanol solution. OPIF may not exist in the brains of day-0 pupae or in brain-subesophageal ganglion and prothoracic ganglion complexes of 5th-instar larvae. The short-day pharate pupae responded to OPIF in a dose-dependent manner.

The small copper butterfly, Lycaena phlaeas daimio, has pupal beige/black polymorphism, the development of which is found to be controlled in an apparent association with the development of adult seasonal polymorphism (spring and summer morphs) by photoperiod and temperature in the larval stages. That is, the pupae of beige and black types developed under long-day and short-day conditions tend to develop into brown-winged and red-winged adults, respectively. In addition, a large proportion of long-day pharate pupae chilled at 4 degrees C for 5 days were observed to develop into pupae whose head-thoracic complexes and abdomens were judged to be of the black and intermediate types, respectively. They developed into adults with redder wings as compared to those obtained from unchilled pupae. The results indicate that the physiological mechanism underlying the photoperiodic control of the development of adult seasonal polymorphism may also play a significant role in the determination of pupal beige/black polymorphism in L. phlaeas daimio. Furthermore, cuticle melanization was found to be induced in the head-thoracic complexes of pupae by chilling of the pharate pupae. Melanization of pupal cuticle seems to occur in a close association with the development of reddish-winged adults.

To understand social organization among Japanese badgers (Meles meles anakuma), we investigated their spatial distribution and sett use in Yamaguchi City, Western Honshu, Japan. Field observations were made from 1995 to 2000, and 19 badgers were monitored by radio-tracking between 1997 and 2000. A badger family consisted of a mother and her offspring. Mature adult males seldom visited the family except in early spring, and they were found to have large home ranges that encompassed the home ranges of two to three adult females. Individual home ranges of mature adult male and adult females did not overlap with those of other mature adult male and other adult females of the same sex, suggesting intra-sexual territoriality. Badgers used an average of 13.5 setts in a year, and mature adult males and females seldom stayed in the same sett together. Male offspring shared a sett with their mother for up to 26 months, whereas female offspring remained with the mother for only 14 months. The average time male offspring spent with their mothers decreased when young were between 15 and 19 months old.

The swallowtail butterfly, Papilio xuthus L., exhibits seasonal diphenism, i.e., spring and summer morphs, the development of which is determined by photoperiod and temperature imposed during the larval stage. Larvae reared under long day conditions at 25°C develop into summer morphs without pupal diapause, while larvae reared under short day conditions at 20°C develop into spring morphs after diapause development at 4°C for more than three months. To investigate the neuroendocrine mechanism regulating seasonal morph development, the activity of the summer-morph-producing hormone (SMPH) was tried to assayed by using male short day pupae of P. xuthus that accomplished diapause development at 4°C. The wing patterns of male butterflies shifted toward summer morphs when short day pupae were treated with the SMPH at an adequate stage. SMPH activity was detected in 2% NaCl extracts of the brain-subesophageal ganglion (Br-SG) complexes of P. xuthus long day pupae. Chilled male short day pupae responded ...

We investigated the neuroendocrine regulation of the development of seasonal morphs in a bivoltine race (Daizo) of the silkmoth, Bombyx mori, by decerebration, the transplantation of brain-suboesophageal ganglion (Br-SG) complexes and the injection of active neuropeptides. When brains were removed from fresh pupae destined to develop into summer morphs (SD pupae) by embryonic and larval exposures to short days at low temperature, the pupae developed into autumn or intermediate morphs. However, in pupae destined to develop into autumn morphs (LD pupae), the operation did not show an effect on seasonal morph development. Br-SG complexes were excised from fifth-instar LD and fifth-instar SD larvae 2 days after larval ecdysis and were transplanted into the abdomen of SD larvae of the same age. The Br-SG complexes of LD larvae, but not the Br-SG complexes of SD larvae, shifted the host's seasonal morph development toward the autumn morph. Furthermore, when treated with crude pupal SGs extract and diapause hormone (DH), fresh SD pupae developed into autumn or intermediate morphs, respectively. Possibly the development of seasonal morphs in the silkmoth, B. mori, is regulated by a novel function of DH. Alternatively, DH may act on the imaginal wing disks at an earlier stage than on the ovaries.

The pupae of Papilio xuthus show green, brown, and orange color polymorphism. The color of the pupal body is determined by a hormone that produces brown coloration, a hormone called pupal-cuticlemelanizing hormone (PCMH) which is secreted from brain-suboesophageal ganglion and prothoracic ganglion (Br-SG-PG) complexes during the pharate pupal stage. PCMH was extracted with 2% NaCl from Br-SG-PG complexes of P. xuthus green pupae. When pharate pupae producing brown pupae (brown-pupa-producers) were ligatured between the thorax and abdomen with cotton thread, the head-thorax complex developed into the brown type and the abdomens into the green type. Ligatured abdomens treated with crude PCMH produced intermediates of green- and brown-pupae (melanization degree of grades 0–3), which was used to assay PCMH-activity. Extracts of Br-SG complexes from Bombyx mori adults also shifted the color of the pupal cuticle toward the brown type in ligatured P. xuthus abdomens of brown-pupa-producers. The molecula...

The cabbage armyworm, Mamestra brassicae, enters winter and aestival diapause in the pupal stage under 10L-14D and 16L-8D, respectively, at 23°C. It develops without pupal dormancy under 14L-10D at 23°C. In 6th-instar larvae, the hemolymph ecdysteroid titers as well as the activity and sensitivity of the prothoracic glands (PGs) were nearly the same in the diapause- and nondiapause-programmed larvae. In non-diapause pupae (ND-pupae), the ecdysteroid titer peaked in the first half of the pupal stage. By contrast, in winter and aestival diapause pupae (WD- and AD-pupae), the ecdysteroid titer remained low after pupation. The ecdysteroid titer in diapausing AD-pupae was 8- to 10-times higher than that in diapausing WD-pupae. PGs from 5th-instar and 6th-instar larvae, as well as those from pupae, secreted ecdysteroids and showed a far stronger response to big prothoracicotropic hormone (PTTH) than to small PTTH in vitro. PG activity and its sensitivity to big PTTH fluctuated in a stage-dependent mann...

A neuropeptide from brain-suboesophageal ganglion (Br–SG) complexes of the silkmoth, Bombyx mori, shows summer-morph-producing hormone (SMPH) activity in the Asian comma butterfly, P. c-aureum. The SMPH-active peptide was extracted and demonstrated to be almost the same molecular size as bombyxin (4–5 kD), a nueropeptide which shows prothoracicotropic hormone (PTTH) activity when assayed in vitro with prothoracic glands (PGs) of 4th-instar B. mori larvae in vitro. A Sephadex G-50 fraction of 3–8 kD molecules prepared from Br–SG complexes of B. mori adults was applied to CM-, SP-, DEAE- or QAE- Toyoperal columns at pH 5.6 (or pH 6.9). The SMPH-activity could be separated from the PTTH-activity (or bombyxin) by subjecting a SMPH- and PTTH-active preparation of B. mori to anion-exchange chromatography at pH 6.9. By reversed-phase HPLC following an anion-exchange chromatography, SMPH-activity was recovered in two fractions of 40–45% acetonitril. Results demonstrate that the B. mori peptide showing the SMPH-activity in P. c-aureum is a different molecule than bombyxin.

To investigate whether four Papilio species, Papilio xuthus, P. machaon, P. bianor and P. helenus, have two molecular forms of the prothoracicotropic hormones (PTTHs), referred to as big- and small-PTTHs, the PTTHs were extracted and fractionated from their pupal brains. The activating ability of big- and small-PTTH fractions was examined by the in vitro assay using the prothoracic glands (PGs) of 2-day-old 5th-instar larvae of their own and several other papilionid species. Big- and small-PTTH fractions activated the larval PGs of their own species to increase the ecdysteroid secretion in vitro. The doses of small-PTTH fractions for activating the larval PGs were 8- to 10-times larger than those of big-PTTH fractions. The big- and small-PTTH fractions as well as those of P. machaon activated the PGs of 2-day-old 5thinstar larvae of several heterogeneous papilionids, but the activating ability did not always decrease with the distance of the genetic (or phylogenetic) relationships. The results in...

Males of the viviparous teleost fish Gambusia affinis copulate with females by using a specialized anal fin, or gonopodium. When female G. affinis were placed in a shallow transparent tank which was then floated on the surface of a larger aquarium housing male G. affinis , the males frequently attempted to copulate with females housed in the smaller quarantine tank. This copulatory behavior in male G. affinis was only observed to be elicited by visual stimuli; female G. affinis have a yellow spot and black anal spots around their urogenital opening. To investigate the function of the yellow spot of the female, we examined male copulatory behavior directed towards artificial female models having a yellow, black, gray, or white spot in the genital region. Of the differently colored females tested, males spent a significantly longer time in the vicinity of the artificial model with a yellow spot. In addition, males attempted to copulate with the yellow-spotted female model more frequently than with the models of different colors. These findings revealed that the yellow spot around the female urogenital opening of G. affinis attracts males and functions as a cue for copulation.

Mutations of the epidermal growth factor receptor (EGFR) gene have been reported in non-small cell lung cancer (NSCLC), especially in female, never smoker patients with adenocarcinoma. Some common somatic mutations in EGFR, including deletion mutations in exon 19 and leucine to arginine substitution at amino acid position 858 (L858R) in exon 21, have been examined for their ability to predict sensitivity to gefitinib or erlotinib. On the other hand, previous report has shown that the insertion mutation at exon 20 is related to gefitinib resistance. We investigated the exon 20 EGFR mutation statuses in 322 surgically treated non-small cell lung cancer cases. Two hundred and five adenocarcinoma cases were included. The presence or absence of EGFR mutations of kinase domains was analyzed by direct sequences. EGFR insertion mutations at exon 20 were found from 7 of 322 (2.17%) lung cancer patients. We also detected the 18 deletion type mutations in exon 19, and 25 L858R type mutations in exon 21. There was a tendency towards higher exon 20 insertion ratio in never smoker (never smoker 4.4% versus smoker 1.3%, p=0.0996) and female (female 4.5% versus male 1.3%, p=0.0917). Two exon 20 insertion cases were treated with gefitinib and failed to response. EGFR insertion mutation in exon 20 could not be ignored from Japanese lung cancers.

Overexpression of the epidermal growth factor receptor (EGFR) is caused by EGFR gene amplification and is sometimes associated with expression of a variant EGFR (deletion exon 2-7 or EGFRvIII). EGFRvIII mutation has oncogenic potential and is investigated as a potential therapeutic target. We genotyped the EGFRvIII mutation status in 252 surgically treated lung cancer cases. The presence or absence of EGFRvIII mutation was analyzed by real-time quantitative polymerase chain reaction (PCR) with mutation specific sensor and anchor probes. EGFR copy number was evaluated with PCR-based assay. EGFR mutation status at kinase domain has been examined and reported. EGFRvIII mutation was found on 8 of 252 patients. All patients were male, smokers, and 7 had squamous cell carcinoma. The mutation status was significantly correlated with pathological subtypes (squamous cell carcinoma vs. adenocarcinoma, p=0.0114). Sixty EGFR mutations at kinase domain exclusively existed with EGFRvIII mutations. EGFR gene copy number was significantly higher in EGFRvIII mutant (4.711+/-4.968) than in non-EGFRvIII mutant (2.284+/-1.224) (p=0.0001). EGFRvIII gene mutation might be one of the mechanisms of increased EGFR copy number. Further studies are needed to confirm the mechanisms of EGFRvIII mutations for possible anti-EGFR therapy for lung cancer.

Several studies have suggested that epidermal growth factor receptor (EGFR) gene mutation, EGFR gene amplification, and some other biomarkers may be predictors of gefitinib sensitivity. We analyzed EGFR mutation and EGFR copy number in 22 gefitinib-treated non-small cell lung cancer (NSCLC) cases and their relation to the survival of patients. We also studied 143 gefitinib-naïve Japanese NSCLC cases. The erbB2 copy number was also studied in 59 gefitinib-naïve NSCLC cases. In gefitinib-treated patients, the presence of EGFR mutation was associated with a higher response rate to gefitinib and a longer overall survival, but the increased EGFR gene copy number was not. In gefitinib-naïve cases, EGFR mutation but not EGFR gene copy number was significantly correlated with gender, pathological subtypes, and smoking status. The erbB2 copy number was not significantly correlated with the EGFR mutation or EGFR copy number in 59 cases. In conclusion, EGFR mutation was a better predictor of clinical outcome in gefitinib-treated patients than the EGFR gene copy number.

We have investigated 26 adenosquamous lung cancer tissues and found that four EGFR mutations were mainly in female and non-smoker lung cancer. However, EGFR mutation at kinase domain was exclusive with K-ras mutation. However, smoking and gender status could affect the occurrence of EGFR mutation. There was no difference in EGFR mutation status if analysis was performed in never smoker female subgroup.

Somatic mutations of the epidermal growth factor receptor (EGFR) gene were found in about 25-40% of Japanese lung cancer patients. These mutations are associated with clinical and radiographic responses to EGFR tyrosine kinase inhibitors. Most common mutation are arginine for leucine substitution at amino acid 858 (L858R) and exon 19 deletions, especially deletion type 1 mutation. We investigated these EGFR mutation statuses in 575 surgically treated non-small cell lung cancer (NSCLC) cases. Three-hundred and sixty-two adenocarcinoma cases were included. The presence or absence of EGFR mutations of kinase domains was analyzed by genotyping analysis (TaqMan assay; n=386, and LightCycler assay; n=98) and sequences (n=91). EGFR mutations (CTG; CGG; L858R) were found from 63 of 575 lung cancer patients. We also detected the deletion 1a type mutations (2235-2249 del GGAATTAAGAGAAGC) from 39 patients and deletion 1b type mutations (2236-2250 del GAATTAAGAGAAGCA) from 15 patients in exon 19. These mutation statuses were significantly correlated with gender, smoking status (never smoker versus smoker), and pathological subtypes (adenocarcinoma versus non-adenocarcinoma). L858R mutation (p

Background The presence of epidermal growth factor receptor (EGFR) mutations in gefitinib-naive lung cancer patients has been reported to be higher in females, in non-smokers, in Japanese, and in adenocarcinoma patients, especially in bronchioloalveolar carcinoma (BAC). To further investigate the prevalence of EGFR mutations in relation to pathological factors, we evaluated EGFR mutations in series of Japanese adenocarcinoma patients who had never been treated with gefitinib. Methods In the previous studies, we examined mutation status in the tyrosine kinase domain of EGFR, exon18 through exon21, in 112 primary lung adenocarcinoma samples. Using these data, adenocarcinomas were histologically classified according to the presence or absence of bronchioloalveolar components. Results Among 112 patients, 48 had adenocarcinoma with BAC components. Those with adenocarcinomas with BAC components had higher frequency of EGFR mutation (28/48, 58%) than those with non-BAC adenocarcinoma (24/64, 37%, P = 0.036). Male patients had the same trend; 12/23 (52%) male patients with adenocarcinoma with BAC components and 10/47 (21%) of those with non-BAC adenocarcinoma had EGFR mutation (P = 0.0135) but there was no correlation between the EGFR mutation status and with/without BAC components in 42 female patients (P = 0.30). Among 11 male non-smokers, patients with adenocarcinoma with BAC components had a tendency to have EGFR mutation more frequently than those with non-BAC adenocarcinoma (P = 0.061). In clear contrast, the frequency of EGFR mutation did not differ significantly between male smoker patients with adenocarcinoma with BAC components and those with non-BAC. Among patients with adenocarcinoma with BAC components, those with adenocarcinoma with EGFR gene mutation had a significantly better 5 year survival than those with adenocarcinoma with wild-type (85.7 versus 46.0%, P = 0.0017). Conclusions Adenocarcinomas with BAC components in male non-smokers seem to predict the presence of EGFR mutation. Half of female adenocarcinoma patients with EGFR mutation exhibit adenocarcinomas with non-BAC suggesting a different behavior from those in males. The prognosis of patients with adenocarcinoma with BAC components with EGFR gene mutation is predicted to be better than that of patients with adenocarcinoma with BAC components with wild-type EGFR gene.

The silken girdles of pupae of the swallowtail butterfly Atrophaneura alcinous show black and white color diphenism. Field observations revealed that all pupae observed on non-food plants and the leaves and stems of the larval food plant Aristolochia debilis were classified as a silken girdle of a black type, while a large portion of pupae pupating on the twigs and trunks of cherry trees in close proximity to A. debilis were classified as a silken girdle of a black type. Additionally, all pupae observed on the surfaces of artificial objects in areas where there are no surrounding plants or trees were classified as a silken girdle of a white type. We demonstrated the effect of day length and the texture, light, plant odor and humidity of pupation sites on the coloration of the silken girdle in A. alcinous. Regardless of long-day or short-day day length conditions, light conditions of constant light or dark, or the presence of a plant odor of A. debilis as environmental cues, all larvae placed at over 80% relative humidity (R.H.) developed into pupae with a silken girdle of a black type. However, all larvae developed into pupae with a silken girdle of a white type when R.H. was below 75%. Furthermore, when pupae with a silken girdle of a white type were transferred to conditions of 90% R.H. within 24 hr of pupation, the white color of the silken girdle changed into a black type within 24 hr of the transfer. The present data suggest that the induction of a black coloration of the silken girdle in A. alcinous requires a R.H. of approximately 80% or more as an environmental factor.

Background Recently, somatic mutations of the epidermal growth factor receptor ( EGFR ) gene and Braf gene were found in patients with lung cancer. These mutations might be correlated with a clinical response to molecular target therapy. Although a few Caucasian lung cancer patients harbored BRAF mutations, there have been no reports about the BRAF mutation in Japanese patients with lung cancer. Materials and methods We investigated BRAF mutations in Japanese lung cancer patients. The study included 129 surgically removed lung cancer cases from Nagoya City University Hospital. Braf , EGFR , and erbB2 mutations also were analyzed by reverse transcript polymerase chain reaction (RT-PCR) and direct sequencing. Results In exon 15, one BRAF mutation (1796 thymine to adenine; V599E) was found in nonsmoking woman with well-differentiated adenocarcinoma. We detected the 43 EGFR mutations, including 19 at exon 19 and 20 at exon 21 from the 129 patients analyzed (33.3%). We also detected one erbB2 mutation from 76 patients analyzed. All these mutations existed exclusively. Conclusions V599E BRAF mutation was uncommon in Japanese lung cancer. All three genes mutations were predominantly found in female nonsmoking subjects with adenocarcinomas. However, completely exclusive mutation status would help us to choose custom-made molecular target therapy for the lung cancer.

Summary Epidermal growth factor receptor ( EGFR ) gene mutations are frequently detected in lung cancer, especially in adenocarcinoma, in females, and non-smoking patients. EGFR mutations are closely associated with clinical response to EGFR tyrosine kinase inhibitor. Bronchioloalveolar carcinoma (BAC) appearance is a good predictor of response to this agent. Noguchi et al. subdivided small peripheral adenocarcinoma of the lung into two groups. One group was characterized with tumor cell growth replacing the normal alveolar cells with varying degree of fibrosis (types A–C), and the other shows non-replacing and destructive growth (types D–F). Using probes for the 13 mutations which have been previously described, we have genotyped the EGFR gene status in surgically resected atypical adenomatous hyperplasias (AAH) and small peripheral adenocarcinomas up to 2cm in diameter using TaqMan PCR assay. In 95 small-sized adenocarcinomas, the EGFR mutations were detected in 37 patients (38.9%), and no mutations were found in five AAHs. In small peripheral adenocarcinomas, EGFR mutations were found 47.1% of types A, B, or C adenocarcinomas; it was less frequent (16%) in Noguchi's types D, E or F adenocarcinomas. These results suggest that type D, F adenocarcinomas are not derived from the less malignant types A–C adenocarcinomas; rather, they have arisen de novo by distinct mechanisms. Although types A and B adenocarcinomas are almost 100% cured by surgery, some type C adenocarcinoma show lymph node metastasis and relapse. EGFR mutation analysis may help identify patients who will respond to treatment with tyrosine kinase inhibitors, e.g., gefitinib.

Purpose: Recently, somatic mutations of the epidermal growth factor receptor (EGFR) gene were found in ∼25% of Japanese lung cancer patients. These EGFR mutations are reported to be correlated with clinical response to gefitinib therapy. However, DNA sequencing using the PCR methods described to date is time-consuming and requires significant quantities of DNA; thus, this existing approach is not suitable for a routine pretherapeutic screening program. Experimental Design: We have genotyped EGFR mutation status in Japanese lung cancer patients, including 102 surgically treated lung cancer cases from Nagoya City University Hospital and 16 gefitinib-treated lung cancer cases from Kinki-chuo Chest Medical Center. The presence or absence of three common EGFR mutations were analyzed by real-time quantitative PCR with mutation-specific sensor and anchor probes. Results: In exon 21, EGFR mutations (CTG → CGG; L858R) were found from 8 of 102 patients from Nagoya and 1 of 16 from Kinki. We also detected the deletion mutations in exon 19 from 7 of 102 patients from Nagoya (all were deletion type 1a) and 4 of 16 patients from Kinki (one was type 1a and three were type 1b). In exon 18, one example of G719S mutation was found from both Nagoya and Kinki. The L858R mutation was significantly correlated with gender (women versus men, P < 0.0001), Brinkman index (600 ≤ versus 600>, P = 0.001), pathologic subtypes (adenocarcinoma versus nonadenocarcinoma, P = 0.007), and differentiation status of the lung cancers (well versus moderately or poorly, P = 0.0439), whereas the deletion mutants were not. EGFR gene status, including the type of EGFR somatic mutation, was correlated with sensitivity to gefitinib therapy. For example, some of our gefitinib-responsive patients had L858R or deletion type 1a mutations. On the other hand, one of our gefitinib-resistant patients had a G719S mutation. Conclusions: Using the LightCycler PCR assay, the EGFR L858R mutation status might correlate with gender, pathologic subtypes, and gefitinib sensitivity of lung cancers. However, further genotyping studies are needed to confirm the mechanisms of EGFR mutations for the sensitivity or resistance of gefitinib therapy for the lung cancer.

A bivoltine race (Daizo) of the silkmoth, Bombyx mori, has two types of adults which are classified on the basis of the number of dark-brown and brown scales distributed along the outer lines of the anterior wings. Development of these two types of adults is determined by photoperiod and temperature experienced during the embryonic and larval stages. The two types of adults are thought to be seasonal (summer and autumn) morphs. All adults developed under short days at low temperature (15 degrees C and 23 degrees ) were classified as summer morphs and oviposited only nondiapause eggs (ND-eggs), and those that developed under long days at high temperature (28 degrees C and 25 degrees C) were autumn morphs ovipositing only diapause eggs (D-eggs), only ND-eggs or a mixture of D- and ND-eggs. Larval exposure to long days at 25 degrees C shifted the wing pattern towards the autumn morphs and decreased the incidence of female adults ovipositing only D-eggs. Larval exposure to long days elicits an opposite effect on D-egg oviposition to that observed when long days are received only during the embryonic stage.

Spring and summer morphs of Lycaena phlaeas daimio Seitz. are characterized by a wing colour of red and reddish brown, respectively. When newly ecdysed pupae destined to be summer or intermediate morphs (90 or 10%) by larval exposure to long days (long-day pupae) were decapitated or decerebrated, more than half of the operated pupae developed into intermediate and spring morphs (48 and 7%). But, in pupae destined to be spring, intermediate, or summer morphs (72, 22 or 4%) (short-day pupae) these operations did not produce any significant changes in the seasonal morph. Brains excised from newly ecdysed long-day pupae were transplanted into the abdomen of decapitated short-day pupae of the same age. The implants changed most recipients into summer and intermediate morphs (46 and 36%). However, when the brains of short-day pupae were used, no significant changes occurred in the seasonal morph. When long-day or short-day pupae were treated with 20-hydroxyecdysone just after pupation, they produced more reddish wings than those of the untreated or saline-treated controls. When the application was followed by chilling, already known to induce the reddish morph, the effects of both treatments are cumulative so that more reddish adults developed. The results indicate that the brain of long-day pupae secretes a factor causing the wing to be brownish. In the absence (or low titre) of the factor, most short-day pupae develop into spring or intermediate morphs. Furthermore, ecdysteroids make the wing more reddish, when applied to newly ecdysed pupae.

In the butterfly, Polygonia c-aureum, development of seasonal forms controlled by the photoperiod and temperature was shown to involve a neuroendocrine system of the brain-corpus cardiacum-corpus allatum complex. For analysis of the neuroendocrine system concerned, the innervation of the complex was investigated first by cobalt chloride perfusion staining and then by severance of axons, ablation of the candidate cells, injection of a homogenate of these cells and transplantation of corpora cardiaca using pupae programmed to be either summer-form or autumn-form adults. The results suggested that medial nerve cells produce what is called material producing the summer form. The seasonal forms of the Asian comma butterfly, Polygonia c-aureum L., summer and autumn forms (Fig. la, b), are determined by the photoperiod and the temperature during the larval period (1–3). Previous studies have given the following results on the physiological mechanism involved in the effect of environmental factors in inducing these seasonal forms. First, the mechanism involves neurosecretory cells located somewhere in the brain (2). Second, the nervous connections between the brain and the corpus cardiacum (NCC I+II (4)) and between the right and left brain lobes are indispensable for the effect (2, 5–7). The present study consisted of two series of experiments. One was designed to demonstrate morphologically the axonal connection of the corpus cardiacum with the corpus allatum in this butterfly, like that shown in several other insects (8–13). The other series was designed to locate the neurosecretory cells producing material related to the seasonal form and to see if this material is also present in the corpus cardiacum.

Seasonal morphs (spring and sumer forms) of Papilio xuthus L. are determined coincidentally with diapause and non-diapause in pupae by larval exposure to short days and long days respectively. The neuroendocrine principle underlying seasonal-morph determination was studied using surgical operations in P. xuthus . When recipient 0-day old or chilled diapause pupae were joined to donor 0-day old non-diapause pupae, the recipients developed into summer or intermediate morphs. When the same kinds of recipients used above were joined to 0-day old or chilled diapause pupae, there were no significant effects on the adult morph. In contrast, recipient non-diapause pupae all developed into summer morphs, regardless of groups of the type of donors. Furthermore, when the brain of 5th-instar larvae, pharate pupae or pupae, predetermined to be diapause, was transplanted into the abdomen of 0-day old, 30-day old or chilled diapause pupae, the recipients developed into summer or intermediate morphs. The results indicate that the brain of non-diapause pupae secretes a humoral factor producing the summer morph. In non-diapause pupae, the factor may be secreted at about the stage of larval-pupal ecdysis coincidentally with that of prothoracicotropic hormone.

To establish the indices evaluating the ecological conditions of forests in Bangladesh, total number of butterflies counted in 5 observation sites reached 27,805 (76 species, 10 families). Number of butterflies (IND) and species showed bimodal pattern of seasonal fluctuations peaked in April and November-to- December. The values of I(R5) also showed bimodal seasonal fluctuations peaked in January-to-February and July-to-September. The ratios of IND/I(R5) formed a large peak in November. Results indicate that species diversities of butterflies high in March-to-July decreased thereafter, the numbers of butterflies increased toward November in the study area. Key words: Butterfly, seasonal fluctuation, population indices, ecological states  doi:10.3329/ujzru.v27i0.1958 Univ. j. zool. Rajshahi Univ. Vol. 27, 2008 pp. 67-71

In Polygonia c-aureum, there are two seasonal forms, viz., the summer form and the autumn form. Previous experiments (Endo, 1970) showed that the corpora allata of the summer form accelerate the maturation of ovarian follicles soon after imaginal ecdysis. On the other hand, the corpora allata of the autumn form do not stimulate ovarian maturation during a period of 2 to 3 weeks after the emergence. In the summer form (S) as well as in the autumn form (A), the corpora allata remained in an inactive state for about 15 days after emergence when they had been isolated microsurgically from the brain and the corpora cardiaca during the larval period. Further, when separation of the brain into the right and left hemispheres or ablation of the medial neurosecretory group cells of the pars intercerebralis had been carried out on S-pupae of 28 hr after pupation or of earlier ages, they developed into autumn form in respect to wing pattern and their corpora allata did not stimulate ovarian maturation until 2 to 3 weeks after emergence. On the other hand, when the above operations had been performed 34 hr after the pupation or of later ages, they developed into summer form in respect to wing pattern and ovarian maturation was evident soon after the emergence. In these cases, stimulation of the corpora allata is closely connected with the development of seasonal-forms of wing pattern. From these experiments, it is clear that the medial neurosecretory group cells of the pars intercerebralis in S-insects stimulate the corpora allata about 30 hr after pupation by way of the nervi corporis cardiaci and the activated corpora allata promote ovarian maturation throughout adult life. In A-insects, on the other hand, the medial neurosecretory group cells of the brain are inactive and fail to activate the corpora allata which in turn have no influence on ovarian maturation.

A field survey of butterflies was carried out in different areas of deciduous forests at Kaliakyar, Gazipur, during January-December, 2008. Seven species of satyrid butterflies were collected and two species, Melanitis leda and Mycalesis mineus were found to exhibit dry and wet season polymorphism. The dry season forms (DSFs) and wet season forms (WSFs) of the both species were distinguished by their upperside and underside wing patterns. The DSFs occurred only in the drier period (November to February) when day length was shorter than 12 hrs. at lower temperatures (24.1-20.6°C). Both the species of butterflies of the wetter period (March - October) were predominant of the WSF when day length was longer than 12 hrs. at higher temperatures (26.8-27.6°C). The fluctuations in the number of seasonal forms of M. leda and M. mineus were higher in DSF than those of WSF. The transition of DSF to WSF and WSF to DSF of M. leda was found to occur around the end of April and the early of September, respectively. But the turnover of DSF to WSF occurred around the early March and the turnover of WSF to DSF in October in M. mineus . Key word: Butterflies; seasons; polymorphism; Melanitis leda; Mycalesis mineus DOI: http://dx.doi.org/10.3329/ujzru.v29i1.9463 UJZRU 2010; 29(1): 33-39

In summer form adults of Polygonia c-aureum, the ovary develops soon after emergence and eggs mature a few days later. However, in autumn form adults, having been kept in laboratory conditions, only several mature eggs are found about four weeks after emergence. The allatectomy during the larval period prevents egg maturation in adults of both forms. When the corpora allata from the summer form of both sexes are transplanted into allatectomized adults of the autumn or summer form, the ovary develops as in the case of summer form adults. But the corpora allata of both sexes of the autumn form are not effective in inducing the ovarian maturation. Environmental factors, especially day-length and temperature during the larval period, may act through the neuroendocrine system to induce the activity of the corpora allata in the adult life.