Comparison of genetic and morphological methods to detect the presence of American lobsters, <Emphasis Type="Italic">Homarus americanus</Emphasis> H. Milne Edwards, 1837 (Astacidea: Nephropidae) in Norwegian waters

American lobsters (Homarus americanus H. Milne Edwards, 1837) are imported live to Europe and should according regulations be kept in land-based tanks until sold. In spite of the strict regulations aimed specifically at preventing the introduction of this species into the NE Atlantic, several specimens of H. americanus have been captured in the wild, especially in Oslofjord, Norway since 1999. One of the great concerns is interbreeding between the introduced American species and the local European lobster, H. gammarus (Linnaeus, 1758). For this reason an awareness campaign was launched in 2000 focusing on morphologically “unusual” lobsters caught in local waters. Morphological characters have been based on colour and sub-ventral spines on the rostrum. Two samples of H. americanus were used for comparisons, as well as samples of European lobster from Oslofjord collected in 1992. Previous genetic analyses (allozymes, mtDNA and microsatellite DNA) have demonstrated that the American lobster is distinct from its European counterpart, with several additional alleles at many loci in addition to different allelic frequency distribution of alleles of “shared” alleles. During the present study, thirteen microsatellite loci were tested in the initial screening, and the three most discriminating loci (Hgam98, Hgam197b and Hgam47b) were used in a detailed comparison between the two species. A total of 45 unusual lobsters were reported captured from Ålesund (west) to Oslofjord (southeast) from 2001 to 2005 and these were analysed for the three microsatellite loci. Nine specimens were identified as American lobsters. Comparisons between morphological and genetic characteristics revealed that morphological differences are not reliable in discrimination the two species, or to identify hybrids. Further, some loci display almost no overlapping in allele frequency distribution for the reference samples analysed, thus providing a reliable tool to identify hybrids.     

Characterization of SSRs from the American lobster,Homarus americanus

The American lobster (Homarus americanus), a commercially important benthic marine crustacean, is widely distributed along the continental shelf of the western North Atlantic. The population substructure of this species remains poorly understood despite its economic value. Informative markers are required to clarify relationships between local populations. To this end, we developed eight polymorphic short sequence repeats (SSR) for the American lobster, which were derived from expressed sequence tags. Additionally, we tested four SSRs previously identified for the Norway lobster (Nephrops norvegicus L.) for cross‐species utility; only one of these showed polymorphism.     

The bronchial tree and blood vessels of the Japanese monkey lung

The author injected various colored celluloid solutions into the bronchial tree and blood vessels of the lungs of five adult Japanese monkeys (Macaca fuscata) in order to prepare cast specimens. These specimens were investigated from the comparative anatomical viewpoint to determine whether the bronchial ramification theory of the mammalian lung (Nakakuki, 1975, 1980) can be applied to the Japanese monkey lung or not. The bronchioles are arranged stereotaxically like those of other mammalian lungs. The four bronchiole systems, dorsal, ventral, medial, and lateral, arise from both bronchi, respectively, although some bronchioles are lacking. In the right lung, the bronchioles form the upper, middle, accessory, and lower lobes, while in the left lung, the upper and accessory lobes are lacking and bi-lobed middle and lower lobes are formed. In the right lung, the upper lobe is formed by the first branch of the dorsal bronchiole system. The middle lobe is the first branch of the lateral bronchiole system. The accessory lobe is the first branch of the ventral bronchiole system. The lower lobe is formed by the remaining bronchioles of the four bronchiole systems. In the left lung, the middle lobe is formed by the first branch of the lateral bronchiole system. The lower lobe is formed by the remaining bronchioles. Thus, the bronchial ramification theory of the mammalian lung applied well to the Japanese monkey lung. The right pulmonary artery runs across the ventral side of the right upper lobe bronchiole. It then runs along the dorso-lateral side of the right bronchus between the dorsal bronchiole system and the lateral bronchiole system. On its way, it gives off branches of the pulmonary artery which run along the dorsal or lateral side of each bronchiole except in the ventral bronchiole system. In the ventral bronchiole system, the branches run along the ventral side of the bronchioles. The distributions of the pulmonary artery in the left lung are the same as those in the right lung. The pulmonary veins do not always run along the bronchioles. Most of them run on the medial or ventral side of the bronchioles. Some of them run between the pulmonary segments. In the right lung, these pulmonary veins finally form the right upper lobe vein, right middle lobe vein and the right lower lobe pulmonary venous trunk before entering the left atrium. However, the right accessory lobe vein runs on the dorsal side of the bronchiole and pours into the right lower lobe pulmonary venous trunk. In four cases out of the five examples, part of the right lower lobe veins pour into the right middle lobe vein, while the others enter the right lower lobe pulmonary venous trunk. In the left lung, the branches of the pulmonary veins finally form the left middle lobe vein and the left lower lobe pulmonary venous trunk.     

Hormonal regulation of fruit set, parthenogenesis induction and fruit expansion in Japanese pear

The effects of applied gibberellins (GAs), GA₁, GA₃, GA₄ and GA₇ with a cytokinin, N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) and indole-3-acetic acid (IAA) on fruit set, parthenogenesis induction and fruit expansion of a number of Rosaceae species were assessed. These included Japanese pear cv. ‘Akibae’ (self-compatible) and cv. ‘Iwate yamanashi’ (a seedless cultivar). Other Rosaceae species (Pyrus communis, Chaenomeles sinensis, Cydonia oblonga, and Malus pumila) were also investigated. GA₄, GA₇ and CPPU are very effective in inducing parthenocarpic fruit growth, whereas GA₁, GA₃ and IAA, have no ability to induce parthenogenesis in Japanese pear. GA₄- and GA₇-induced parthenocarpic fruit tended to be smaller in size, higher in flesh hardness, and showed advanced fruit ripening in comparison to pollinated fruit and to parthenocarpic fruit induced by CPPU. GA₄- and GA₇-induced parthenocarpic fruit also had an increased pedicel length and fruit shape index and also showed a slight protrusion of the calyx end. CPPU, GA₄ and GA₇ alone or combination with uniconazole were also active in inducing parthenogenesis in three other Rosaceae species, although final fruit set was extremely low. GA₁ was essentially inactive in promoting fruit expansion unlike the other bioactive GAs, whose effectiveness in promoting fruit cell expansion was as follow: GA₄ ≈ GA₇ > GA₃ > GA₁.     

Morphology and distribution of antennular setae of scyllarid lobsters (Scyllarides aequinoctialis, S. latus, and S. nodifer) with comments on their possible function

Abstract. Lateral flagella of the antennules of scyllarid lobsters were examined for setal morphology and distribution via scanning electron microscopy. Setal distribution patterns were mapped directly for 3 regions of the antennule ( base, tuft , and tip ) and analyzed for differences: (1) between left and right antennules, (2) between males and females within a species, and (3) among species by comparing counts of setae per annulus in the ventral tuft region only. Six types of antennular setae were identified based on their external morphology: aesthetases, simple, modified simple, asymmetric, hemi-plumose, and toothbrush setae. These different types were organized in a clear pattern over the ventral and dorsal surfaces of the lateral flagella of the antennule. Aesthetase, asymmetric, modified simple, and hemi-plumose setae were found only on annuli in the tuft region between the distal and proximal ends of the flagellum. Simple setae were found on all annuli of all regions of the antennule, and toothbrush setae were mainly concentrated on all annuli of the base region and on proximal annuli of the tuft region . All species of scyllarids examined had the same general pattern of setal distribution and no differences were found between left and right, or male and female antennules. Similar setae located on the lateral antennules of species from the families Nephrophidae and Palinuridae (clawed and spiny lobsters) have been previously described as chemo- and/or mechanoreceptive for use in distance chemoreception (i.e., detection and orientation to olfactory stimuli). Based on work on clawed and spiny lobsters, we predict that the aesthetases on slipper lobsters have a chemoreceptive function and that simple and toothbrush setae may have a bimodal chemo- and mechanoreceptive function.     

Circadian rhythms of heart rate in freely moving and restrained American lobsters, Homarus americanus

While circadian rhythms of locomotion have been reported in the American lobster, Homarus americanus, it is unclear whether heart rate is also modulated on a circadian basis. To address this issue, both heart rate and locomotor activity were continuously monitored in light-dark (LD) cycles and constant darkness (DD). Lobsters in running wheels exhibited significant nocturnal increases in locomotor activity and heart rates during LD, and these measures were significantly correlated. In DD, most lobsters exhibited persistent circadian rhythms of both locomotion and heart rate. When heart rate was monitored in restrained lobsters in LD and DD, most animals also demonstrated clear daily and circadian rhythms in heart rate. Overall, this is the first demonstration of circadian rhythms of heart rate in H. americanus, the expression of which does not appear to be dependent on the expression of locomotor activity.     

Circadian rhythms of heart rate in freely moving and restrained American lobsters,Homarus americanus

While circadian rhythms of locomotion have been reported in the American lobster, Homarus americanus, it is unclear whether heart rate is also modulated on a circadian basis. To address this issue, both heart rate and locomotor activity were continuously monitored in light-dark (LD) cycles and constant darkness (DD). Lobsters in running wheels exhibited significant nocturnal increases in locomotor activity and heart rates during LD, and these measures were significantly correlated. In DD, most lobsters exhibited persistent circadian rhythms of both locomotion and heart rate. When heart rate was monitored in restrained lobsters in LD and DD, most animals also demonstrated clear daily and circadian rhythms in heart rate. Overall, this is the first demonstration of circadian rhythms of heart rate in H. americanus, the expression of which does not appear to be dependent on the expression of locomotor activity.     

Secular trend and age distribution of cribra orbitalia in Japanese

Cribra orbitalia is currently regarded as an indicator of environmental conditions. The prevalence and severity of this lesion in Japanese populations from prehistoric Jomon to modern times were recorded and discussed. Since the lesion was low in both frequency and grade in the late and final Jomon people from Chiba Prefecture, they presumably lived in relatively favorable environmental conditions. The increased prevalence of the lesion in the medieval Muromachi citizens of Kamakura may be attributed to the poor hygiene and social conditions. The high prevalence and marked severity of the lesion in the citizens of Edo (the old name of Tokyo) in the 17th century suggested that they lived under extremely unfavorable situations of nutrition, sanitation and medical care. Cribra orbitalia was most prevalent in adolescence (81.3%) among the Edo citizens. The incidence of the most severe type of lesion rose gradually during childhood, reached a peak in adolescence, and then gradually decreased with age. These findings would reflect intense marrow hyperplasia in the orbital roof associated with both iron deficiency anemia of long standing during the immature period and increased iron requirement in adolescence. The results obtained support the hypothesis that iron deficiency anemia is the primary factor in the etiology of cribra orbitalia. It was concluded that the incidence and severity of cribra orbitalia due to various anemic reactions reflected the dietary, hygienic, social and environmental circumstances of the populations investigated.     

The control of cardiac rhythmicity and of blood distribution in crustaceans

Heart beat arises myogenically in primitive crustaceans, but neurogenic dive is superimposed in malacostracans. The vascular system of malacostracans possesses extensive arterialization whereas those of the more primitive classes do not. These two new innovations (neurogenicity and arteries) offer functional advantages to the malacostracans in terms of neural and hormonal control of the heart and vasculature, flexibility of hemodynamics, and of hemolymph distribution. These advantages may have allowed the evolutionary explosion in the malacostracans in terms of animal size and speciation. The nervous and hormonal controls of isopod and decapod cardiac function are analogous to the control system in vertebrates. The distribution of hemolymph in isopods and decapods is controlled in part by cardioarterial valves, which are in turn under nervous and hormonal control. In decapods the peripheral resistance of arteries is also differentially controlled by hormones and neurotransmitters.     

Blood lead level distribution by by age group in Japanese

Blood lead levels of 634 healthy Japanese (422 males and 212 females), aged 0–87, living in the Kyushu and Okinawa Islands, were determined by the colorimetric dithizone method and atomic absorption spectrometry. The lead level in the atmospheric environment was not extremely high; it was below 1 μg/m³ in most of the places examined. The differences in the geometric means of blood lead levels between 0–5 yr and other age groups were statistically significant (P<0.01). The blood lead levels of Japanese seem to increase in late teens and then decrease slowly to a steady state. The accumulation of lead taken from food was calculated by assuming the amount of lead from food corresponds to the caloric intake recommended for each Japanese age group (e.g., assuming 150 μg/2700 kcal for boys in late teens). Lead concentration in the air was assumed to be 1 μg/m³. The pattern of lead accumulation in blood by age was similar to the theoretical curve for the accumulated total amount of lead obtained by simulation. The fact probably indicates that blood lead levels in Japanese are dependent on lead in daily foods rather than on lead in the atmospheric environment.     

Pleistocene glaciation events shape genetic structure across the range of the American lobster, Homarus americanus

A north/south discontinuity along the northeastern coast of North America in the genetic structure of the American lobster ( Homarus americanus ) was detected using a suite of 13 microsatellite loci assessed using spatial analyses. Population genetic data laid over existing data on physiographic changes and sea-surface temperatures were used to reconstruct the Pleistocene distribution of this species. A postglacial northern-edge colonization model best explains the relative genetic homogeneity of the northern region compared to the southern region centred in the Gulf of Maine. Population genetic analyses identified significant structure (range of standardized theta 0–0.02) but no significant evidence for isolation by distance. The novel application of spatial genetic analyses to a marine species allowed us to interpret these results by providing a greater insight into the evolutionary factors responsible for shaping the genetic structure of this species throughout is natural range.     

Neural mechanisms governing distribution of cardiac output in an isopod crustacean,Bathynomus doederleini: reflexes controlling the cardioarterial valves

In Bathynomus doederleini all of the cardioarterial valves located at the origin of the lateral arteries are dilated by impulses of lateral cardiac nerves. Tactile stimuli applied to sensillar setae depress impulse activities of the 1st and 5th lateral cardiac nerves. The 1st lateral cardiac nerve controls the valve of the lateral artery which runs to the walking-legs and viscera. The 5th lateral cardiac nerve controls the valve of the lateral artery which runs to the swimmeret muscles. The response indicates that tactile receptor reflexes bring about decreased haemolymph flow to the organs. Augmented swimmeret movements were always accompanied by an increased firing rate in the 5th lateral cardiac nerve. Artificial full protraction of swimmerets simultaneously induced excitation of the 5th lateral cardiac nerve and inhibition of the 1st lateral cardiac nerve. The excitation corresponds to an increase in haemolymph flow to the swimmerets, and the inhibition a decrease in haemolymph flow to walking-legs and viscera. Three kinds of mechanoproprioceptors which were activated by swimmeret movements were found. Two of the mechanoproprioceptors are located at the base of the basipodite. The other mechanoproprioceptor supplies processes to a nerve to the retractor muscles. Activation of three kinds of mechanoproprioceptors, induced by artificial swimmeret protraction, triggered lateral cardiac nerve reflex responses.Abbreviations LA lateral artery - LCN lateral cardiac nerve - RMN nerve to retractor muscles - StR stretch receptor     

Low-density lipoprotein receptor-related protein 5 variant A1330V is a determinant of blood pressure in Japanese males

We examined the influence of the A1330V variant in the low-density lipoprotein receptor-related protein 5 gene on blood pressure in a large cohort of Japanese workers. This study used analysis of covariance in a multivariate general linear model to adjust for other potential factors such as age, body mass index, blood chemistry and lifestyle. The target subjects were 1440 males and 1169 females selected from 3834 male and 2591 female workers in a single company. Hypertension was defined as systolic blood pressure >or=140 mm Hg and/or diastolic blood pressure >or=90 mm Hg or the use of antihypertensive medications. Genotype distributions for A1330V in hypertensive males (AA=139(54.5%), AV=101(39.6%), VV=15(5.9%)) and females (AA=48(63.2%), AV=24(31.6%), VV=4(5.3%)) were not significantly different from normotensive males (AA=594(50.1%), AV=488(41.2%), VV=103(8.7%)) and females (AA=568(52.0%), AV=441(40.3%), VV=84(7.7%)). Allele distributions were not significantly different in either sex. In males, analysis of covariance showed that the VV genotype was associated with a 2.5 mm Hg lower diastolic blood pressure and a 2.3 mm Hg lower mean blood pressure than the AA genotype. This study indicates that the 1330V allele is an independent factor for lower diastolic and mean blood pressure in Japanese males.     

Change in the telomere length distribution with age in the Japanese population

Telomeres play a role in cellular aging and they may also contribute to the genetic basis of human aging and longevity. A gradual loss of the telomeric repeat sequences has been reported in adult tissue specimens. This study determined the percentage of telomere restriction fragment in various molecular-sized regions in addition to measuring the average telomere length. Mean telomere restriction fragment (TRF) length was determined by Southern blot analysis using a longer telomeric repeat probe with higher sensitivity. A significant decrease in longer telomere fragments and a quick increase in the shortest fragments were observed, especially in male subjects. There was a tendency that the age-adjusted telomere length was longer in females than that observed in males, while males lose the telomeric sequence faster than females. These data indicated that the percentage of longer telomeres fragments decreased, while the shortest fragments increased quickly with age. In addition, the longest telomere fragments decreased and the short fragments increased with a relatively stable frequency with age. There was also a significant difference in the longest telomere fragment percentage between males and female in their 40s and 50s, whereas no difference was observed in the mean TRF length. Interestingly, the changing rate of the longest and the shortest range group of TRF percentage associated with aging seemed quite different between before and after 50-year old with a gender-related contrast. This contrast implies a drastic change around the age of 50 of unknown factors that affect telomere attrition. The first two authors equally contributed to this article.     

Glutathione S-transferases in the Japanese quail: Tissue distribution and purification of the liver isozymes

Cytosolic glutathione S-transferase (GST) activities toward 1-chloro-2,4-dinitro-benzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), ethacrynic acid (EA), 1,2-epoxy-3-(p-nitrophenoxyl)propane (EPNP), trans-4-phenyl-3-buten-2-one (t-PBO), δ³-androstene-3,17-dione (ASD) and trans-stilbene oxide (t-SO); cytosolic glutathione peroxidase activity toward cumene hydroperoxide (CuOOH); and microsomal GST activity toward CDNB were examined in liver, kidney, brain, and lung of adult male and female Japanese quail. In all cases, the renal specific activity per milligram protein was higher than the hepatic activity and was the highest among the four tissues examined. No consistent sex differences in GST activity were observed. The GSTs were purified from quail liver cytosol by S-hexylglutathione and glutathione affinity chromatography. Total GSTs eluted from the S-hexylglutathione affinity column were further separated by chromatofocusing, and the microheterogeneity of the GST isozymes was shown by high-resolution native isoelectrofocusing (IEF) in polyacrylamide slab gels and by SDS-PAGE. Five subunits were identified: QL1 (30.5 kDa), QL2 (27.2 kDa), QL3a (26.8 kDa), QL3b (26.5 kDa), and QL4 (25.5 kDa). Western blot analysis revealed that QL1 and QL2 reacted with antibodies raised against the rat Mu class GSTs (Yb1 and Yb2), and QL3a and QL3b reacted with those raised against the Alpha class (rat Ya and mouse a). Substrate specific activity of each isoform was determined with CDNB, DCNB, CuOOH, EA, t-PBO, ASD, and t-SO. QL3a and QL3b have high reactivity toward CuOOH, while QL1 and QL2 showed high activity toward t-SO. The N-terminal amino acid sequence of QL2 was identical to that of the chicken Mu class GST subunit CL2. However, no sequence was obtained with QL1 due to possible N-terminal blockage. © 1996 John Wiley & Sons, Inc.     

379株血培养病原菌的临床分布及耐药性分析

目的了解血培养病原菌种类、临床分布及耐药情况。方法对2008年6月至2009年6月的379株血培养病原菌及其药敏情况用WHONET5.4软件进行统计分析。结果 379株病原菌中,分离率从高到低依次是凝固酶阴性葡萄球菌(133/35.1%)、大肠埃希菌(44/11.6%)、肺炎克雷伯菌(32/8.4%)、鲍曼不动杆菌(30/7.9%)、铜绿假单胞菌(25/6.6%)、金黄色葡萄球菌(18/4.7%)、肠球菌(13/3.4%)和其他(75/22.2%)。这些菌株主要分布在ICU和普外科。药敏结果分析显示:革兰阳性球菌中,未发现利奈唑胺和万古霉素耐药株。金黄色葡萄球菌和凝固酶阴性葡萄球菌对苯唑西林的耐药率分别为61.1%和89.5%;屎肠球菌对多数抗生素耐药率超过80%,粪肠球菌对青霉素类抗生素敏感性较高。革兰阴性杆菌中,大肠埃希菌和肺炎克雷伯菌中产ESBLs菌株比例分别为36.4%和31.3%,且发现2株耐亚胺培南的肺炎克雷伯菌;鲍曼不动杆菌除对卡那霉素保持敏感外,对其他抗生素的耐药率为50%~100%;铜绿假单胞菌对头孢曲松的耐药率为100%,对其他β-内酰胺类抗生素、氨基糖苷类和喹诺酮类抗生素的耐药率相对较低。结论临床上应规范血培养标本留取方法以减少污染,加强细菌耐药监测、严格抗生素使用,以延缓细菌耐药情况的发生和发展。     

外科血培养标本中病原菌分布及耐药性分析

目的了解中山大学附属第一医院外科血标本中病原菌的菌种分布及常见菌株的耐药性。方法血标本用Bact/A lert-120全自动血培养仪进行血培养,阳性血培养转种后用VITEK-60 AMS细菌鉴定仪鉴定,用K-B法进行药敏试验。结果2002年1月至2005年12月血培养标本中共分离出病原菌256株,阳性率为10.6%。122株(47.7%)为革兰阴性杆菌,其中肠杆菌科细菌占72.1%(88/122),非发酵菌占27.9%(34/122);113株(44.1%)为革兰阳性球菌,其中葡萄球菌属占51.3%(58/113),肠球菌占38.1%(43/113);真菌21株(8.2%)。血培养中的革兰阴性杆菌对亚胺培南、美洛培南治疗敏感;革兰阳性球菌对万古霉素和替考拉宁敏感。结论肠杆菌科细菌和葡萄球菌是外科血培养中的主要病原菌,其耐药现象严重,宜根据药敏结果选用敏感抗菌药物治疗。     

Cortical bone remodeling rates in a sample of African American and European American descent groups from the American Midwest: comparisons of age and sex in ribs

This study employs regression analysis to explore population and sex differences in the pattern of age-associated bone loss, as reflected by histomorphometric variables that are measures of intracortical and endocortical bone remodeling. A comparison of an African American sample from the Washington Park Cemetery in St. Louis, Missouri, and a European American rib sample composed of cadavers, autopsies, and forensic cases from Missouri reveals the existence of complex age-associated patterns for differences in measures of intracortical remodeling and cortical area. Females from the two samples express similar bone dimensions and dynamics. The African American females appear to lose more bone than their male counterparts, but this difference is absent in the European American sample. When age-associated patterns are considered, it is in the younger cohorts that African Americans exhibit greater relative cortical area than European Americans, but this is reversed in the older ages, when the latter group manifests greater bone mass. The European American males consistently differ in the slopes and intercepts for the variables compared to the other groups, and differences are highly significant with African American females, with the former group maintaining bone mass while the latter exhibit a more rapid bone loss. Achieving larger relative cortical area due to smaller endosteal area, coupled with better bone quality due to lower intracortical porosity early in life, may be a mechanism by which African Americans, especially females, maintain adequate bone mass in older ages, which buffers them from bone loss and related fragility fractures despite higher rates of intracortical remodeling and endosteal expansion later in life. These results suggest that both genetic and environmental factors are responsible for the differences in bone remodeling and bone mass observed between these samples.