Observation of microorganism colonies using a scanning-laser-beam pH-sensing microscope

The extracellular pH-distribution of colonies of Saccharomyces cerevisiae (yeast) and Escherichia coli (E. coli) were observed using a newly-developed scanning-laser-beam pH-sensing microscope. Colonies were incubated either on top of agarose plates or between the pH-sensing surface and the agar. In the latter case, colony growth was observed in-situ. The colonies could be observed within a period as short as 8 h for E. coli. The pH-distribution profiles by the colonies were found to be very sharp, in agreement with simulation results.     

Encystment of the ciliate Boveria teredinidi in the tissues of the molluscan woodborer Bankia gouldi in Barnegat Bay,New Jersey

Histological sections of Bankia gouldi from several stations in Barnegat Bay, New Jersey, were found to contain encysted Boveria teredinidi. The process of encapsulation appeared to be of the leukocytic type, and there was usually considerable leukocytic infiltration into the tissues in the areas of the cysts. No cysts were observed in tissues of Teredo navalis, T. furcifera, or T. bartschi, although ciliates were observed on the gill surfaces of these species. This report is believed to be the first of encystment in B. gouldi.     

Phosphoglucose isomerase polymorphism in the shrimp Metapenaeus affinis

Three phenotypes of phosphoglucose isomerase (PGI) were detected in abdominal muscle extracts from the shrimp (Metapenaeus affinis) by starch-gel electrophoresis. The observed phenotypes were assumed to be under the control of two allelic genes. This assumption was supported by the observed distribution of phenotypes. There were no significant differences in the distribution of PGI phenotypes among samples of shrimp taken from Basrah waters, Iraq and from Kuwaiti waters, Arab Gulf. The three PGI alleles observed in M. affinis appear to be the same in the two localities.     

Shell structure of two polar pelagic molluscs, Arctic Limacina helicina and Antarctic Limacina helicina antarctica forma antarctica

The purpose of this study was to investigate shell growth performance in two thin-shelled pelagic gastropods from cold seawater habitats. The shells of Arctic Limacina helicina and Antarctic Limacina helicina antarctica forma antarctica are very thin, approximately 2–9 μm for shells of 0.5–6 mm in diameter. Many axial ribbed growth lines were observed on the surface of the shell of both Limacina species. Distinct axial ribs were observed on the outermost whorl, while weak or no rib-like structures were observed on the inner whorls in the larger shell of L. helicina antarctica forma antarctica. For L. helicina, no ribs were observed on small individuals with three whorls, while larger individuals had distinct ribs on the outer whorls. Shell microstructure was examined in both species. There is an inner crossed-lamellar and extremely thin outer prismatic layer in small individuals of both species, and a distinct thick inner prismatic layer was observed beneath the crossed-lamellar layer in large Antarctic individuals. Various orientations of the crossed-lamellar structure were observed in one individual. Shell structure appeared to be different between the Antarctic and Arctic species and among shells of different size.     

Phosphoglucose isomerase polymorphism in the freshwater crab Sesarma boulengeri

Three phenotypes of phosphoglucose isomerase (PGI) were detected in abdominal muscle extracts from freshwater crab (Sesarma boulengeri) by starch-gel electrophoresis. The observed phenotypes were assumed to be under the control of two allelic genes. This assumption was supported by the observed distribution of phenotypes. There were no significant differences in PGI phenotype distribution among samples of crab taken from Karmat Ali, and Abu Al-Khasib locations along Shatt Al-Arab River. The three PGI alleles observed in S. boulengeri appear to be the same in specimens from the two locations.     

Multiple-Antibiotic Resistance of Enterococcus spp. Isolated from Commercial Poultry Production Environments

The potential impact of food animals in the production environment on the bacterial population as a result of antimicrobial drug use for growth enhancement continues to be a cause for concern. Enterococci from 82 farms within a poultry production region on the eastern seaboard were isolated to establish a baseline of susceptibility profiles for a number of antimicrobials used in production as well as clinical environments. Of the 541 isolates recovered, Enterococcus faecalis (53%) and E. faecium (31%) were the predominant species, while multiresistant antimicrobial phenotypes were observed among all species. The prevalence of resistance among isolates of E. faecalis was comparatively higher among lincosamide, macrolide, and tetracycline antimicrobials, while isolates of E. faecium were observed to be more frequently resistant to fluoroquinolones and penicillins. Notably, 63% of the E. faecium isolates were resistant to the streptogramin quinupristin-dalfopristin, while high-level gentamicin resistance was observed only among the E. faecalis population, of which 7% of the isolates were resistant. The primary observations are that enterococci can be frequently isolated from the poultry production environment and can be multiresistant to antimicrobials used in human medicine. The high frequency with which resistant enterococci are isolated from this environment suggests that these organisms might be useful as sentinels to monitor the development of resistance resulting from the usage of antimicrobial agents in animal production.     

Nucleoside diphosphate kinase-1 regulates hyphal development via the transcriptional regulation of catalase in Neurospora crassa

A nucleoside diphosphate kinase-1-disrupted (ndk-1^RIP-1) mutant was observed to be defective in aerial hyphal and conidial development. In this study, two types of hyphae, fine and thick, were observed in wild-type (Wt) strains. However, only fine-type hyphae were observed in the ndk-1^RIP-1 mutants. The ndk-1^RIP-1 mutants were stimulated by oxidative stress and constitutively expressed an antioxidant enzyme catalase (CAT)-3. Furthermore the ndk-1^RIP-1 mutants could form thick hyphae by oxidative stress and a disruption of cat-3. These results suggest that the loss of thick hyphae in the ndk-1^RIP-1 mutants may be caused by the over-expression of cat-3.     

Some Physical Characteristics of the Enzymes of l-Tryptophan Biosynthesis in Higher Plants

Anthranilate synthetase, phosphoribosyltransferase, phosphoribosyl anthranilate isomerase, and indoleglycerol phosphate synthetase were examined in partially purified extracts of the monocotyledon, Zea mays and the dicotyledon, Pisum sativum. The plant extracts were chromatographed on DEAE-cellulose and Sephadex G150. The molecular weights of the enzymes were determined and found to be similar to those observed for many bacteria. None of the plant tryptophan enzyme activities was aggregated in vitro as is also the case with most bacteria. This is in contrast with the complex aggregation patterns observed in other eucaryotic organisms that have been examined (fungi and Euglena gracilis). The tryptophan enzymes from peas and corn were generally similar but some differences in stability were observed.     

Aspects of the life cycle of Schistosoma margrebowiei infection in laboratory mammals

Schistosoma margrebowiei Le Roux, 1933 has been recorded in a number of mammals in Africa. The parasite was maintained in the laboratory using Bulinus natalensis as intermediate host and hamsters, mice and gerbils as definitive hosts.Worm recovery, growth of paired worms, egg output and egg viability were determined and compared in the three laboratory hosts. The results in the three hosts are discussed and the mouse was observed to be suitable for the long term studies on S. margrebowiei in the laboratory. Hamsters and gerbils were observed to be useful for the study of the acute phase of the disease.     

Effect of heavy metals (Cu,Cd and Pb) on aspartate and alanine aminotransferase in Ruditapes philippinarum (Mollusca: Bivalvia)

The accumulation of cadmium, copper and lead and their effects on aspartate and alanine aminotransferases in digestive gland, gills, foot and soft body in the clam Ruditapes philippinarum were examined. The animals were exposed to different concentrations: Cd (200–600 μg·l⁻¹), Pb (350–700 μg·l⁻¹) and Cu (10–20 μg·l⁻¹) for 7 days. The highest concentrations were found in digestive gland for cadmium and copper, and in gills for lead, and the lowest values were observed in the foot. Aspartate aminotransferase activity (AST), in general, was not inhibited by cadmium, lead or copper during the exposure. Only in clams exposed to cadmium (600 μg·l⁻¹, 7 days) and copper (20 μg·l⁻¹, 5 days) were observed significant differences (P<0.05) in foot and gills, respectively, with respect to control. In the case of alanine aminotransferase activity (ALT), significant differences were observed for cadmium and lead in treated animals with respect to control. With regard to copper, a decrease in ALT was observed in gills and foot exposed to 20 μg·l⁻¹. A significant correlation (P<0.05) was observed between ALT and metal accumulation for cadmium, copper and lead in gills. In the case of soft body, only cadmium and lead showed a significant correlation. In summary, R. philippinarum can be considered a bioindicator species for cadmium and lead accumulation and ALT could be useful as biomarker of sublethal stress for these metals in soft tissues and gills. Only gills can be considered an adequate target tissue for copper.     

A Comparison of Low-Temperature and Ambient-Temperature SEM for Viewing Nematode Faces

Faces of lesion nematodes Pratylenchus teres (populations RTB and JK) and P. zeae or the bacterivore Distolabrellus veechi were observed on frozen specimens with low-temperature scanning electron microscopy and as chemically fixed, critical-point dried specimens with conventional scanning electron microscopy. Amphidial secretions were preserved in chemically fixed but not cryofixed lesion nematodes. Overhanging liplets of chemically fixed D. veechi may be artifactual because they appeared as variably filled, mostly empty membranes when cryofixed. The diagnostically useful lips of the frozen lesion nematodes exhibited six sectors of variable prominence that were absent in chemically fixed specimens. This variability may be due to different degrees of muscle contraction captured during cryofixation, which occurs in milliseconds. This is the first evidence that rarely observed lip sectors in Pratylenchus may be something other than an artifact of shrinkage.     

Circadian rhythm of homocysteine is hCLOCK genotype dependent

Homocysteine (Hcy) is known to be a prognostic marker for neurological, cardiovascular and cerebrovascular diseases and several other pathophysiological conditions. A sudden surge in Hcy can increase cardiovascular events. Hemodynamic modulations are known to be associated with individual’s chronotype. Therefore, precise monitoring of Hcy is crucial for evaluating its impact on risk. The aim of the present study was to investigate the rhythmicity of Hcy under controlled dietary conditions and whether this rhythmicity is under the genetic control of circadian rhythm. Five subjects were selected from 200 Malayalam speaking healthy ethnic individuals who were screened for functionally critical variants of MTHFR and hCLOCK genes. MTHFR is the rate-limiting enzyme in the methionine cycle and critical for regulating Hcy levels while hCLOCK is a critical gene responsible in regulating the day and night cycles. Rhythmicity in Hcy levels were observed in all the subjects with a consensus on a morning nadir and an evening peak. Gender specific stratification of Hcy levels were observed among similar genotypes of MTHFR and hCLOCK genes. Variations from the conventional rhythmicity of Hcy were observed among similar genotypes of MTHFR and dissimilar hCLOCK genotypes. A reduced plasma Hcy in hCLOCK rs1801260 CC genotype individuals were observed in contrast to CT genotype individuals. The study tends to suggest that Hcy and body time are genetically interdependent and throws light on some of the previously unexplained reasons for variability in Hcy levels. A population specific variation of MTHFR and hCLOCK genes also highlights ethnicity specific risk management.     

Survival and growth of alpha-spores of Porphyra schizophylla Hollenberg (Rhodophyta: Bangiophyceae)

Alpha-spores of the intertidal Porphyra schizophylla Hollenberg f. norrisii Mumford appear able to tolerate environmental stress factors associated with intertidal habitats, (viz., drying of water films with salt crystal formation, increased salt concentrations in the surrounding water, exposure to humid air when spores are enclosed in water films), and to produce Conchocelis filaments of normal appearance when subsequently germinated in sea water cultures. Tolerances of lowered salinities were observed down to 17.4%., but little survival at salinities lower than this. Alpha-spore germination on thalli of intertidal macro-algae was observed, with initial penetration in some cases. Penetrations of algal thalli were observed with Conchocelis filaments of α-spores germinated on red algae from low intertidal and subtidal habitats. Cell recognition phenomena would seem to be involved in these growth responses.     

Energy status of cells lacking dystrophin: an in vivo/in vitro study of mdx mouse skeletal muscle

Although great strides have been made in understanding the genetics of Duchenne muscular dystrophy (DMD), uncertainty still remains as to the metabolic changes which are associated with the disease. We have used the recently discovered animal model of DMD, the mdx mouse, to study aspects of high energy phosphate metabolism and metabiolic control indices in dystrophic muscle. This model of DMD has the dual advantage of having a genetic defect which is homologous to that in human DMD, and it lacks the fatty infiltration and ncecrosis which makes biochemical analysis of DMD so difficult. We have used nuclear magnetic resonance sperctroscopy (NMR) to monitor developmental changes in high energy phosphates and pH. No differences were observed between young (< 40–50 days old) control and mdx mice. The pH increase and alterations in phosphate ratios (i.e., decline in PCr/ATP) observed in adult mdx vs. control mice are quantilatively similar to those observed in humans. Biochemical analysis showed a small decline in ATP and PCr content and a decline in some indices of energy status in adult mdx mice. As young mdx mice appeared to be normal, the lack of dystrophin does not correlate with metabolic changes. The changes which were observed were small enough that alterations in fibre composition could be the major contributory factor.     

Orbital tumors in USA: Difference in survival patterns

Introduction: There is a wide range of tumors affecting the orbital adnexa. Key such tumors include lymphomas, carcinomas, melanomas and rhabdomyosarcomas. Several studies have proposed that these histological subtypes differ in their survival outcomes. In this study we aim to describe the difference in survival outcomes between such subtypes. Methods: The SEER database was used to gather patient information. All 18 SEER registries were used. Patients diagnosed from 1996 to 2005 were included in the analysis. Observed five-year survival rate was calculated using the SEER*Stat software version 8.1.2. Data were extracted into IBM SPSS version 20 to generate Kaplan Meier curve for each group. Results: There were 2180 patients in the SEER databases who met the selection criteria. Lymphomas were the most common histology in adults. The overall five-year observed survival for all lymphoma patients was 75.9% (95% CI: 73.7–78.1). There was statistically significant difference between observed survival rates of lymphoma subtypes. Carcinomas were the second most common tumors. Their five-year observed survival rate in our study was 60.4%. There was no statistically significant difference between carcinoma subtypes’ observed survival rates in the 20–49 age group, while, in the older age group, the difference was found to be statistically significant. Rhabdomyosarcomas were the most common tumors in children. The overall five-year observed survival rate for rhabdomyosarcomas patients was 89.8%. There was no statistically significant difference between observed survival rates of rhabdomyosarcomas subtypes. There was no statistically significant difference between relative survival rates according to gender and treatment received except within melanomas. Conclusion: In adults, lymphomas have better survival rates than carcinomas. Whereas the lymphoma subtype can be used as a determinant prognostic factor in any age, the carcinoma subtype can be used as such a determinant in older age groups only. In children, rhabdomyosarcomas are the predominant tumors affecting the orbital adnexa. Further studies are needed to determine if the difference between embryonal rhabdomyosarcoma and alveolar rhabdomyosarcoma observed survival rates are statistically significant.     

Leishmania adleri: in vitro phagocytosis by lizard leukocytes and peritoneal exudate cells

The interaction of iguanid lizard (Dipsosaurus dorsalis) blood cells and promastigote forms of Leishmania adleri Heisch 1954 was observed in vitro by means of wet mount preparations. Some parasites were phagocytized within a short time after their addition to the preparations. Parasites were engaged and engulfed either individually or in masses. Rapid decrease in size of the parasites occurred. The process of phagocytosis of the parasites was found to be very similar to that of mammalian leishmanias in similar preparations with mammalian cells. Cell cultures prepared from peritoneal exudates of lizards (Basiliscus vittatus) were inoculated with L. adleri promastigotes. Resultant intracellular amastigotes were observed up to 36 hr at 25 and 35 C.     

Rhizobial infection in Adesmia bicolor (Fabaceae) roots

The native legume Adesmia bicolor shows nitrogen fixation efficiency via symbiosis with soil rhizobia. The infection mechanism by means of which rhizobia infect their roots has not been fully elucidated to date. Therefore, the purpose of the present study was to identify the infection mechanism in Adesmia bicolor roots. To this end, inoculated roots were processed following conventional methods as part of our root anatomy study, and the shape and distribution of root nodules were analyzed as well. Neither root hairs nor infection threads were observed in the root system, whereas infection sites—later forming nodules—were observed in the longitudinal sections. Nodules were found to form between the main root and the lateral roots. It can be concluded that in Adesmia bicolor, a bacterial crack entry infection mechanism prevails and that such mechanism could be an adaptive strategy of this species which is typical of arid environments.     

Comorbid Gastric Adenocarcinoma and Gastric and Duodenal Strongyloides stercoralis Infection: A Case Report

Strongyloides stercoralis can cause systemic infection, termed strongyloidiasis, and gastrointestinal ulcer disease in immunocompromised patients. However, to our knowledge, there are no reported cases of comorbid gastric adenocarcinoma and S. stercoralis infection. Here, we report a case of an 81-year-old Korean man who presented with S. stercoralis infection coexisting with early gastric adenocarcinoma (T1aN0M0). S. stercoralis eggs, rhabditiform larvae, and adult females were observed in normal gastric and duodenal crypts. They were also observed in atypical glands representative of adenocarcinoma and adenoma. Preliminary laboratory tests revealed mild neutrophilic and eosinophilic leukocytosis. A routine stool test failed to detect rhabditiform larvae in the patient’s fecal sample; however, S. stercoralis was identified by PCR amplification and 18S rRNA sequencing using genomic DNA extracted from formalin-fixed paraffin-embedded tissues. Postoperatively, the patient had a persistent fever and was treated with albendazole for 7 days, which alleviated the fever. The patient was followed-up by monitoring and laboratory testing for 4 months postoperatively, and no abnormalities were observed thus far. The fact that S. stercoralis infection may be fatal in immunocompromised patients should be kept in mind when assessing high-risk patients.     

Cultivation of avian malaria parasites in mammalian liver cells

The exoerythrocytic stages of Plasmodium lophurae and P. fallax were grown in cell cultures derived from embryonic mouse livers. Liver cell monolayers were maintained in continuous culture with frequent subculturing for extended periods of time. Morphologically, the main cell type was a large, flat cell which closely resembled in shape the mouse parenchymal cell, although small colonies of spindle-shaped cells could also be found. Mammalian cells were labelled with thymidine-³H prior to infection with avian merozoites so they could be positively distinguished from any avian cells which might be present in the merozoite inoculum as contaminants. Merozoites were observed inside the labelled mammalian cells within three hours and large forms were seen at 48 hr. The mammalian cells supported parasite growth comparable to that observed in avian cell cultures.