bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of an endogenous nitric oxide synthase inhibitor on phorbol myristate acetate‐induced acute lung injury in rats
Authors: Lin Hen I , Chu Shi Jye , Wang David , Chen Hsing I , Hsu Kang
Published: July 2003

1. In the present study, we determined whether the endogenous nitric oxide (NO) synthase (NOS) inhibitor Nω‐nitro‐l‐arginine methyl ester (l‐NAME) could ameliorate the acute lung injury (ALI) induced by phorbol myristate acetate (PMA) in rat isolated lung.

2. Typical ALI was induced successfully by PMA during 60 min of observation. At 2 µg/kg, PMA elicited a significant increase in microvascular permeability (measured using the capillary filtration coefficient Kfc), lung weight gain, lung weight/bodyweight ratio, pulmonary arterial pressure (PAP) and protein concentration of bronchoalveolar lavage fluid.

3. Pretreatment with the NOS inhibitor l‐NAME (5 mmol/L) significantly attenuated ALI. None of the parameters reflective of lung injury showed significant increase, except for PAP (P < 0.001). The addition of l‐arginine (4 mmol/L) blocked the protective effective of l‐NAME. Pretreatment with l‐arginine exacerbated PMA‐induced lung injury.

4. These data suggest that l‐NAME significantly ameliorates ALI induced by PMA in rats, indicating that endogenous NO plays a key role in the development of lung oedema in PMA‐induced lung injury.
Evaluation of Withania somnifera in a middle cerebral artery occlusion model of stroke in rats
Authors: Chaudhary Geeta , Sharma Uma , Jagannathan Naranamangalam R , Gupta Yogendra K
Published: July 2003

1. Stroke causes brain injury in millions of people worldwide each year. Despite the enormity of the problem, there is currently no approved therapy that can reduce infarct size or neurological disability. One of the approaches that can be used in limiting the neurological damage after stroke is the use of prophylactic treatment in patients with a high‐risk of stroke. The present study was undertaken to investigate the effect of the Indian herbal plant Withania somnifera as a prophylactic treatment in the middle cerebral artery (MCA) occlusion model of stroke in rats.

2. Two groups of male Wistar rats were pretreated with a hydroalcoholic extract of W. somnifera (1 g/kg, p.o.) for 15 and 30 days. Thereafter, rats were subjected to focal ischaemia by occlusion of the MCA using an intraluminal thread. After 2 h MCA occlusion, reperfusion was allowed by retracting the thread. Animals were assessed for ischaemic changes using diffusion‐weighted imaging 30 min after reperfusion. Twenty‐four hours later, rats were subjected to motor performance tests and were subsequently killed for the estimation of the marker of oxidative stress malondialdehyde (MDA). The control group received vehicle and a similar protocol was followed.

3. Significant motor impairment, with elevated levels of MDA, was observed in vehicle‐treated MCA‐occluded rats. In addition, diffusion‐weighted imaging showed increased signal intensity in the right hemisphere compared with the contralateral hemisphere. Treatment with W. somnifera for 15 days did not improve motor performance or decrease the elevated levels of MDA. However, when the pretreatment time of W. somnifera was increased to 30 days, it prevented motor impairment and significantly decreased the raised levels of MDA compared with vehicle‐treated rats. In the W. somnifera (30 days)‐pretreated group, the percentage hemispheric lesion area in diffusion‐weighted imaging was significantly attenuated (17 ± 2%) compared with the vehicle‐treated MCA‐occluded group (30 ± 4%).

4. Because W. somnifera has been documented to have anti‐oxidant properties, the protection afforded by W. somnifera could be due to its anti‐oxidant effect. The present study provides first evidence of the effectiveness of an Indian herb in focal ischaemia.
Improvement of vascular function by chronic administration of a cyclo‐oxygenase inhibitor in cholesterol‐fed rabbits
Authors: Srisawat Supath , Phivthong‐ngam Laddawal , Unchern Supeenun , Chantharaksri Udom , Govitrapong Piyarat , Sanvarinda Yupin
Published: July 2003

1. Atherosclerotic cardio‐ and cerebrovascular disease is a leading cause of mortality in Western countries. Aspirin‐like drugs are widely used to prevent and treat these occlusive cardio‐ and cerebrovascular diseases. The beneficial effects of these drugs have been largely attributed to inhibition of platelet cyclo‐oxygenase activity and thromboxane (TX) A2 production. We investigated the effect of an aspirin‐like drug, namely indomethacin, on endothelial function, plaque and platelet aggregation and the formation of vasoactive substances during the development of atherosclerosis in cholesterol‐fed rabbits.

2. Rabbits were fed 1% cholesterol (n = 8), 1% cholesterol plus 25 mg/day indomethacin (n = 8) or normal rabbit chow (control group; n = 8) for 12 weeks. Urinary excretion rates of 2,3‐dinor‐TXB2, 6‐keto‐prostaglandin (PG) F1α, 8‐iso‐PGF2α and nitrate were analysed at the beginning of dietary intervention and at 4 weekly intervals thereafter. At the end of the study period, platelet aggregation, aortic plaque formation and endothelium‐dependent and ‐independent vascular functions of isolated aortic rings ex vivo were assessed.

3. Compared with control, in the cholesterol‐fed group, urinary 2,3‐dinor‐TXB2, 6‐keto‐PGF1α and 8‐iso‐PGF2α excretion and platelet aggregation were significantly increased (P < 0.05), but urinary excretion of nitrate was decreased (P < 0.05). Treatment with indomethacin significantly reduced platelet aggregation, urinary 2,3‐dinor‐TXB2, 6‐keto‐PGF1α and 8‐iso‐PGF2α excretion (P < 0.05 vs the cholesterol‐fed group) and attenuated the reduction in urinary nitrate excretion.

4. Cholesterol feeding progressively increased aortic intimal thickening and impaired endothelium‐dependent vasodilator function (P < 0.05 vs control), whereas indomethacin partially prevented aortic plaque formation and restored endothelium‐dependent vasodilation (P < 0.05 vs the cholesterol‐fed group).

5. The present study demonstrates that indomethacin reduces the progression of atherosclerotic lesions and improves endothelium‐mediated vascular responses ex vivo in cholesterol‐fed rabbits. The beneficial effects of indomethacin may be due to its ability to prevent the elevation of platelet aggregation, TXA2 (measured as urinary 2,3‐dinor‐TXB2 excretion) and 8‐iso‐PGF2α formation and to retard the decrease in endogenous nitric oxide synthesis (assessed as urinary excretion of nitrate). Despite indomethacin treatment leading to the suppression of prostacyclin biosynthesis (assessed as urinary 6‐keto‐PGF1α excretion), according to our data, indomethacin appears to preserve endothelial function.
Function of renal angiotensin AT2 receptors is not enhanced in Lyon hypertensive rats
Authors: Liu Kiao Ling , Lo Ming , Benzoni Daniel , Sassard Jean
Published: July 2003

1. Because we previously observed that angiotensin AT2 receptor stimulation decreased pressure–natriuresis, in the present study we examined the possible involvement of these receptors in altered sodium excretion shown by Lyon hypertensive (LH) rats.

2. In 9‐week‐old male anaesthetized LH rats and normotensive (LL) controls pretreated with an angiotensin‐converting enzyme inhibitor (quinapril; 10 mg/kg) and an AT1 receptor antagonist (losartan; 10 mg/kg), angiotensin (Ang) II was infused (30 ng/kg per min) to stimulate AT2 receptors. In other groups of rats, an AT2 receptor antagonist (PD123319; 50 µg/kg per min) was added before AngII infusion.

3. During AT2 receptor stimulation, LH differed from LL rats by significantly reduced renal blood flow (RBF), glomerular filtration rate and pressure diuresis and natriuresis. The addition of PD123319 did not change total RBF, whereas it did increase pressure diuresis and natriuresis in both strains. However, the effects of PD123319 were less marked in LH rats than in LL rats.

4. These findings confirm that, under the present experimental conditions, AT2 receptors are antinatriuretic and are not of greater functional importance in hypertensive animals of the Lyon strain.
Changes in autonomic activity and baroreflex sensitivity with the hypertension process and age in rats
Authors: Nagai Ryuji , Nagata Shinya , Fukuya Fumiyo , Higaki Jitsuo , Rakugi Hiromi , Ogihara Toshio
Published: July 2003

1. Autonomic activity and baroreflex sensitivity (BRS) were compared in age‐matched conscious groups of Wistar Kyoto (WKY) rats, spontaneously hypertensive rats (SHR) and stroke‐prone spontaneously hypertensive rats (SHRSP).

2. Male WKY rats, SHR and SHRSP aged 4–30 weeks were used. Autonomic activity and BRS were estimated by power spectral and cross‐spectral analysis of systolic blood pressure (SBP) and SBP–SBP (SS) interval fluctuations, respectively.

3. The time‐course of heart rate (HR), SBP, the amplitude of the low‐frequency component of SBP fluctuation (SBP‐LF; prazosin‐sensitive index) and the amplitude of the high‐frequency component of the SS interval fluctuation (SS‐HF; atropine‐sensitive index) consisted of two periods. In the first period (up to 10 or 15 weeks of age), BP, SBP‐LF and SS‐HF increased with age. The order of SBP‐LF was SHRSP > SHR > WKY rats throughout this period. During the second period, BP was sustained at certain levels in all strains, but changes in SBP‐LF and SS‐HF with age were different among strains. In particular, in SHRSP, SBP‐LF markedly decreased with age after 10 weeks. Baroreflex sensitivity in WKY rats increased gradually with age, whereas the BRS in SHR and SHRSP decreased before 6 weeks of age and remained lower than that in WKY rats.

4. In conclusion, the present study shows that both prazosin‐sensitive and atropine‐sensitive indices are associated with the elevation of BP in all strains studied. However, hypertension after 15 weeks of age in SHRSP is sustained despite a paradoxical reduction in sympathetic activity with an abnormal control of BRS. Therefore, the contribution of autonomic activity to hypertension may be discussed separately as a developmental period and a sustained period.
Role of mitochondrial KATP channels and protein kinase C in ischaemic preconditioning
Authors: Uchiyama Yuka , Otani Hajime , Wakeno Mayu , Okada Takayuki , Uchiyama Takamichi , Sumida Tomohiko , Kido Masakuni , Imamura Hiroji , Nakao Shinichi , Shingu Koh
Published: July 2003

1. Activation of mitochondrial KATP (mitoKATP) channels and protein kinase C (PKC) has been implicated in cardioprotective mechanisms of ischaemic preconditioning (IPC). However, the exact role of these events in early IPC remains unclear.

2. Isolated and perfused rat hearts underwent IPC with three cycles of 5 min ischaemia and 5 min reperfusion. The heart was subjected to 30 min global ischaemia followed by 120 min reperfusion. Flavoprotein oxidation was monitored to assess mitoKATP channel activity. Cardioprotection was evaluated by recovery of isovolumic left ventricular (LV) function and infarct size.

3. Diazoxide (50 mgr;mol/L) increased flavoprotein oxidation and conferred cardioprotection in a manner sensitive to the selective mitoKATP channel blocker 5‐hydroxydecanoate (5‐HD; 0.5 mmol/L).

4. Pretreatment with 0.5 mmol/L 5‐HD abrogated IPC‐induced flavoprotein oxidation and cardioprotection, whereas late treatment with 5‐HD after IPC required a higher dose (2 mmol/L) to abolish flavoprotein oxidation and cardioprotection afforded by IPC.

5. Pretreatment with the PKC inhibitors Ro318425 (1 µmol/L) and chelerythrine (5 µmol/L) abolished IPC‐induced flavoprotein oxidation and cardioprotection, whereas late treatment with Ro318425 required a higher dose (4 µmol/L) to abolish flavoprotein oxidation and cardioprotection.

6. In conclusion, these results suggest that activation of mitoKATP channels is the trigger and the mediator of IPC and that PKC plays a crucial role in both phases of mitoKATP channel activation, although mitoKATP channels and PKC may be more activated during the mediator phase.
Lateral and vertical crown associations in mixed forests
Authors: AKASHI NOBUHIRO , KOHYAMA TAKASHI , MATSUI KIYOSHI
Published: July 2003

The crown‐association sampling method was developed to examine horizontal and vertical spatial associations in forest tree communities. We sampled tree crowns along line transects and recorded the lateral contact of neighboring crowns within the canopy and the vertical overlap of crowns between the understory and the canopy. Deviations from random association can be examined for lateral and vertical associations. It is also possible to compare the results for vertical overlap among study sites. Three natural mixed forests were sampled: a warm‐temperate rainforest on Yakushima Island (southern Japan), and two cool‐temperate mixed forests in Ohdaigahara (central Japan) and on the Shiretoko Peninsula (northern Japan).
Role of the diurnal variation of urinary pH and urinary calcium in urolithiasis: A study in outpatients
Authors: Murayama Tetsuo , Sakai Naoki , Yamada Tetsuo , Takano Tetsuzou
Published: July 2003

Abstract Background: We previously reported that the urinary excretion of calcium and the diurnal variation of urinary pH were important for stone formation in hospitalized inpatients with hospital standard diet. Because almost all urinary stones are formed in outpatients with ambulatory free diet, it is essential to investigate these factors in outpatients.

Methods: We examined the urinary excretion of calcium, oxalate, uric acid, phosphate and magnesium in 96 male outpatients and 142 male inpatients. We also studied the relationship between the stone composition and the diurnal variation of urinary pH in 32 male outpatients (five uric acid stones (UA), 13 pure calcium oxalate stones (CaOX) and 14 mixed calcium oxalate and calcium phosphate stones (CaOX–CaP)) and 53 male inpatients (nine UA, 15 CaOX and 29 CaOX–CaP).

Results: There was a significant difference in the urinary excretion of calcium among outpatients with UA, CaOX and CaOX–CaP (133 ± 96 vs 219 ± 97 vs 268 ± 102 mg per day, P < 0.05). In outpatients with UA, urinary pH was constantly low throughout the entire day. In contrast, outpatients with CaOX and those with CaOX–CaP had diurnal variation of urinary pH that was low in the early morning, followed by elevation in the daytime and was lowered in the night.

The pHs in the early morning, afternoon and night were significantly higher in outpatients with CaOX–CaP than in those with CaOX.

Conclusion: The diurnal variation of urinary pH and the urinary calcium are important for stone formation.
Comparison of Japanese General Rules of Prostatic Cancer and Gleason grading system
Authors: Fukagai Takashi , Namiki Thomas , Carlile Robert G. , Sugawara So , Morita Masashi , Shimada Makoto , Yoshida Hideki , Namiki Hideo
Published: July 2003

Abstract Background: We compared the prognostic efficacy between the Japanese General Rules of Prostatic Cancer (JGRPC) and the Gleason grading system (GGS) by applying them to a single set of patients and assessing the survival outcome.

Methods: One hundred and seventy‐six patients with previously untreated prostate cancer were studied. One experienced Japanese pathologist graded the slides with JGRPC. Another experienced American pathologist graded the same slides with the Gleason grading system. The JGRPC grades were correlated with the Gleason scores (GS) grouped into three (GS 2–4, 5–7 and 8–10) or four (GS 2–4, 5–6, 7 and 8–10) tiers.

Results: The highest cancer death rates were seen in the higher grade groups in both systems. Comparison of JGRPC grade and three‐tiered grouping of the GS showed identical grades in 81 of 176 cases (46.0%). The overall kappa value of agreement was only 0.151. The 96 cases of JGRPC moderately differentiated carcinoma group contained two nearly equal‐sized groups by the Gleason grading system, those with GS 5–7 (47cases) and GS 8–10 (49 cases). There was a significant difference in survival rate between the GS 5–7 and GS 8–10 groups. No significant differences were noted in the reverse analysis of survival by JGRPC groups within patients with the same GS three‐tiered groups. Similar trends were seen when JGRPC was compared with the four‐tiered grouping of the GS.

Conclusion: Both JGRPC and the Gleason grading system are useful in estimating the prognosis of prostate cancer, but only a mild correlation was found between the two systems. The Gleason grading system may provide more prognostic information than JGRPC in the moderately differentiated group.
The Landscape Game: A learning tool demonstrating landscape design principles
Authors: McIntyre By S.
Published: July 2003

Summary A board game was developed for use in workshops to demonstrate some theoretical principles underpinning conservation planning at the landscape scale. The game is based on neutral landscape models and demonstrates the effects of habitat removal and arrangement on landscape connectivity for organisms with different mobility characteristics. This paper briefly describes the ecological principles underpinning the game, the equipment required to run a game session, and the rules of the game. The game was piloted in three workshops where it was played by a total of 75 people, primarily extension personnel. It proved to be an effective learning tool that was adopted by a number of the participants for their own communication activities.
The role of small ground‐foraging mammals in topsoil health and biodiversity: Implications to management and restoration
Authors: Martin By Greg
Published: July 2003

Summary Many early Australian records indicate that at the time of European settlement there were extensive tracts of highly productive, and species‐rich, grassy communities and chenopod shrublands. Topsoils in many areas were soft and friable. The rapid development of livestock industries led to most changes to the environment being simplistically ascribed to domestic stock grazing, land clearance, introduced pests (such as rabbits) or changed burning practices. It has also commonly been assumed that the hoof action of domestic stock was the principal cause of the compaction and surface sealing of soils in many areas. However, the rapid soil deterioration also coincided with the dramatic decline or complete extinction of many small native ground‐foraging mammals and the consequent cessation of the soil disturbances and interactions that they created. This paper reviews the role of small mammals in disturbing soils, and implications for incorporation of organic matter, aeration, improvement in infiltration and the provision of suitable sites for seed germination and seedling establishment. This can aid topsoil formation and health by providing substrate for microorganisms, improved water balance and mineral cycles and enhanced soil structure. Seeds and mycorrhizal fungi, that are integral to the establishment and growth of many plants, are spread. Such intermittent disturbance may be an important driving force in determining the pathway of succession and lead to greater biodiversity. Further ongoing research on Australia's small mammals is needed, especially in areas where they are able to move freely in a natural environment and are protected from introduced predators.
Buffel Grass and fire in a Gidgee and Brigalow woodland: A case study from central Queensland
Authors: Butler By Don W. , Fairfax Russell J.
Published: July 2003

Summary The effects of a fire on plant community structure were examined in a remnant Gidgee (Acacia cambagei) and Brigalow (Acacia harpophylla) woodland infested with the naturalized exotic, Buffel Grass (Cenchrus ciliaris) in central Queensland. Fifteen months after the fire, burnt areas had about half the basal area of living trees and more than twice the cover of Buffel Grass and Parthenium Weed (Parthenium hysterophorus) as unburnt areas. This is consistent with the idea that Buffel Grass invasion, which increases ground fuel loads in Acacia woodlands, is facilitated by burning, producing positive feedback between Buffel Grass and fire. The result is accelerating remnant degradation, making the interaction between Buffel Grass and fire an important target for management measures. Fire‐breaks and fuel reduction strategies including periodic intense grazing and canopy enhancement have potential to reduce the impact of Buffel Grass invasion, but long‐term community survival will probably require effective control of Buffel Grass. The impact of weed invasion within remnant vegetation is clearly a complex issue but the simple ‘case study’ approach employed here can both increase and communicate understanding.
The impact of grazing by Eastern Grey Kangaroos (Macropus giganteus) on vegetation recovery after fire at Reef Hills Regional Park, Victoria
Authors: Meers By Trevor , Adams Robyn
Published: July 2003

Summary In southeastern Australia ecological burning is frequently used to maintain a number of plant and animal populations. However, many of these prescribed fires are small, and may focus intense grazing activity on new regrowth. At Reef Hills Regional Park, Victoria shrub species have senesced, presumably due to the absence of fire. Ecological burning may be necessary to promote regeneration, however, the population density of the Eastern Grey Kangaroo (Macropus giganteus) is high (approx 38 per km2), and grazing pressure presents a significant risk to postfire vegetation recovery. An assessment of grazing patterns and their effects on postfire recovery was carried out at Reef Hills Regional Park through grazing exclusion plots. Preferential grazing by Eastern Grey Kangaroos occurred on small burnt plots compared to adjacent unburnt areas as determined by faecal pellet counts. On burnt areas, there was a significant reduction in shrub diversity on grazed plots compared to ungrazed plots. Most observations of kangaroos were of animals grazing on farmland surrounding the Park, and it is likely that any burning might shift grazing from farmland to burnt areas when new growth occurs. This needs to be considered before any ecological burn plan is applied to manage vegetation communities, particularly if the plan requires small areas to be burnt. We recommended that a large area up to 200 ha area be burnt and monitored to determine whether burning larger areas disperses grazing pressure from macropods to a level where impacts on vegetation are reduced and localized plant extinctions do not occur.
The effect of concentrated smoke products on the restoration of highly disturbed mineral sands in southeast Victoria
Authors: Coates By Terry D.
Published: July 2003

Summary Recent studies have recognized the potential of broad‐scale surface application of smoke compounds for enhancing germination from the soil seed‐bank in fire‐prone vegetation communities. Results suggest that smoke technology may play, in the future, a significant role in the restoration and management of areas supporting indigenous vegetation. An important step in the development of smoke‐based restoration tools is the conduct of in situ field trials in a range of geographical locations and environmental conditions. However, most of the published work on the effectiveness of smoke products in promoting seedbank germination has been conducted at sites in southwestern Australia. The present study examines the effect of commercially available smoke‐water products on the regeneration of a highly disturbed former mine‐site at the Royal Botanic Gardens Cranbourne, in southeastern Victoria. Various combinations of concentrated smoke products and topsoil harvested from a nearby heathy woodland community were applied to exposed, uniform mineral sands to test their effect on seedling density and species richness of regrowth. The trials showed that after 12 months a number of common, herbaceous species including Austrodanthonia setacea, Opercularia varia and Platysace heterophylla were recorded in significantly higher numbers in areas treated with a commercial smoke‐water. However, there was no overall improvement in the density of seedlings or the richness of species as a result of the application of the smoke products. Similarly, total seedling density and species richness were not affected by the addition of topsoil, either alone or in combination with smoke products.
Endoscopic Diagnosis and Treatment of Local Residual/Recurrent Lesions after Endoscopic Mucosal Resection for Early Colorectal Carcinoma
Authors: TANAKA SHINJI , OKA SHIRO , NAGATA SHINJI , ITO MASANORI , HARUMA KEN , CHAYAMA KAZUAKI
Published: July 2003

Local residual/recurrent lesions have been observed with some frequency after endoscopic mucosal resection (EMR) for colorectal tumors. Many reports have revealed that the rate of recurrence after piecemeal resection is higher than that after en bloc resection. Thus, to accomplish an appropriate trimming in EMR, it is important to closely observe the lesions to be resected, possibly by magnification. Our data show that, with careful trimming of lesions, there are no significant differences in rates of local residual recurrence between en bloc and piecemeal resections. The manner of recurrence and the biological characteristics of residual/recurrent tumors depend on whether the resected lesion is an adenoma, carcinoma in adenoma, de novo carcinoma, mucosal (m) or submucosal (sm) carcinoma. Therefore, it is essential to choose the appropriate method of follow‐up observation according to histopathologic findings of resected lesions. In local residual/recurrent lesions of intramucosal carcinomas, the treatment policy should be decided from an overall evaluation of histological findings on both recurrent and resected primary lesions. After EMR of sm carcinomas, attention should always be paid to both the loci of resection and possible metastasis during follow‐up observation; surgical treatment is inevitable in the case of recurrence.
Indications and Techniques for Endoscopic Mucosal Resection in the Lesions of A Colorectal Tumor
Authors: TAMURA SATORU , OHKAWAUCHI KOJI , YOKOYAMA YUICHI , ONISHI SABURO
Published: July 2003

We evaluated the possibility of an extended application of endoscopic treatment for submucosal‐invading colorectal cancers, and describe the method of endoscopic mucosal resection (EMR) using a one channel colonoscope.

A total of 328 submucosal‐invading cancers were examined from July 1985 to September 2002. The patterns of infiltrating growth into the submucosal layer were further divided into two groups: expanding growth, and infiltrating growth. Lymph node metastasis occurred in sm2 and extension cancer in more advanced stages. The lowest measurement of submucosal invading cancer with lymph node metastasis was 1250 µm. As for patterns of invasion, the frequencies of lymph node metastasis in the groups of expanding growth and infiltrating growth were 0% (0/87) and 14.5% (16/110), respectively, (P = 0.0002, Fisher's direct method). Results showed that endoscopic treatment is suitable for sm1 extension without vessel invasion, but there is a possibility that some sm2 extension cancers can be cured radically when the pattern of submucosal invasion shows expanding growth with a distinct border.
Newly Developed Endoscopic Resection Technique for Colorectal Tumors
Authors: CHO EISAI , UNO KOJI , TANAKA KIYOHITO , YASUDA KENJIRO , NAKAJIMA MASATSUGU
Published: July 2003

We applied a newly developed endoscopic resection technique for a rectal mucosal cancer of 4.2 cm. This method resulted in a curable treatment and provided precise information for histological examinations. This technique, using IT‐knife, may involve the risk of bleeding and perforation compared with conventional methods. Further improvements are needed to make this technique safer and more reliable for a standard endoscopic method for large colorectal tumors.
Present Situation of Eus‐Fnab in our Institute and Future Appropriate Direction of This Procedure in Japan
Authors: HIROOKA YOSHIKI , ITOH AKIHIRO , HASHIMOTO SENJU , NIWA YASUMASA , GOTO HIDEMI
Published: July 2003

We describe the present view of performing diagnostic endoscopic ultrasonography guided fine needle aspiration biopsy (EUS‐FNAB). In the pancreatic diseases, our application of EUS‐FNAB for pancreatic disease has been as follows: as to the operable cases, EUS‐FNAB should not be performed through any pathway penetrating the duodenum and the stomach. EUS‐FNAB is contraindicated in cystic diseases. As to the cases diagnosed as inoperable with various imaging modalities, EUS‐FNAB should be performed to obtain the pathological evidence. In gastrointestinal disorders, all intramural diseases are appropriate indications for this procedure. Even if the target lesion is in the far oral part of the large intestine, we perform EUS‐FNAB by the special method described in this paper. Lymphadenopathy for appropriate cases is one of the best cases in which to perform EUS‐FNAB. Mediastinal lymph node metastasis influences the treatment plan of lung cancer; the application of this treatment is increasing in our institute.
The Results of Questionnaire about Endoscopic Mucosal Resection in the Stomach
Authors: KATO MOTOTSUGU , SHIMIZU YUICHI , NAKAGAWA SOUICHI , SUGIYAMA TOSHIRO , ASAKA MASAHIRO
Published: July 2003

Isolated exfoliation method of gastric endoscopic mucosal resection (EMR) as a new technique has not yet reached the popularity of the conventional EMR techniques. From the results of a questionnaire about EMR in the stomach, the isolated exfoliation method has the advantage of permitting en bloc and histologically complete resection regardless of lesion size. However, this method has the disadvantage of long performance time and high frequency of complication as well as the need for a high level of technical skill. New devices and ideas are needed for the development of the isolated exfoliation method.
Endoscopic Mucosal Resection for Early Gastric Cancer: Aiming at Safety, Speed, and Reliability
Authors: FUJISAKI JUNKO , MATSUDA KOJI , TAJIRI HISAO
Published: July 2003

We assessed the non‐marginating surgical techniques of endoscopic mucosal resection (EMR) for early gastric cancer, represented by the ‘double‐channel’ method and the suction method, and the marginating surgical techniques, represented by the ‘insulation tip’ (IT) and the ‘endoscopic resection with high saline epinephrine’ (ERHSE) methods, and reviewed cases in the literature and our own cases from the standpoint of complications and duration of therapy with each method. Bleeding occurred in 3.6–4.5% of the cases treated by the non‐marginating surgical techniques and 17% of the cases treated by the marginating surgical techniques. Perforation occurred in only 0.3% of the cases treated by the non‐marginating surgical techniques, as opposed to 6.7% of the cases treated by the marginating surgical techniques. The results of EMR with a new type of therapeutic scope, the multi‐bending scope, showed that it was useful for obtaining a direct en face view of the lesser curvature of the body of the stomach, and an increase in the rate of one‐time complete resection was seen when it was used.
Prospect of Endoscopic Mucosal Resection for Early Gastric Cancer: Our Devices in Insulated‐Tip Electrosurgical Knife Method
Authors: MORITA YOSHINORI , AOYAMA NOBUO , SHIRASAKA DAISUKE , FUKUDA MASATERU , KASUGA MASATO
Published: July 2003

In Kobe University Hospital, a new method for endoscopic mucosal resection (EMR) using insulated‐tip electrosurgical knife (IT‐EMR) for early gastric cancer (EGC) was introduced from November 2001. To achieve an effective and safe IT‐EMR procedure, we use a high‐frequency surgical unit for cutting and coagulation (ERBOTOM ICC 200) with automatically controlled cutting mode (ENDOCUT). In this study, we show not only our results of IT‐EMR for EGC in comparison with those of the conventional strip biopsy method, but also the optimal conditions for the apparatus of a high‐frequency surgical unit to prevent complications such as bleeding and perforation.
Enodscopic Mucosal Resection for Gastric Cancer: Necessity of ‘Incision and Stripping Method’ and Present Status
Authors: KIDA MITSUHIRO , TANABE SATOSHI , SAIGENJI KATSUNORI
Published: July 2003

Endoscopic mucosal resection (EMR) was developed in the early 1980s and has been employed widely as a radical treatment for certain groups of early gastrointestinal cancer because of its lower invasiveness, cost effectiveness, and short hospital stay. In the late 1990s extension of its indication has been discussed and several data allow us to perform EMR in wide lesions more than with conventional indications. However, using conventional EMR such as the 2‐channel method, cap method, etc. it becomes difficult to remove lesions more than 2 cm en bloc. Then ‘incision and stripping’, such as insulated‐tipped diathermic knife (IT knife), needle knife with doom food, flex‐knife, and hook knife were also developed. Using these techniques, it becomes feasible to remove larger lesions more than 2 cm en‐bloc. However, in order to improve these techniques it is necessary to perform a large number of the procedures.
Theoretical and Technical Requirements to Expand Emr Indications
Authors: YAHAGI NAOHISA , FUJISHIRO MITSUHIRO , IGUCHI MIKITAKA , KAKUSHIMA NAOMI , OMATA MASAO
Published: July 2003

A large study of surgically resected gastric cancers revealed several conditions of mucosal cancers without lymph node metastasis, and if we are able to resect such lesions in an en bloc fashion, it would be judged to be a curative treatment. With the use of submucosal dissection method, giant lesions, e.g. over 10 cm that occupy the entire lesser curvature of the stomach, lesions of difficult locations, e.g. the esophago‐gastric junction and lesions with submucosal fibrosis caused by ulcer scars, have been treated successfully. These technical progresses have enabled the application of endoscopic mucosal resection to many lesions that meet theoretical conditions.
Endoscopic Mucosal Resection of Early Esophageal Cancers: Our Method
Authors: PARK SEUNG WOO , PARK YOUNG SOO , SONG SI YOUNG
Published: July 2003

Though endoscopic mucosal resection (EMR) may provide definitive treatment for selected cases of early esophageal carcinoma, the standard method has not been determined yet. There have been several EMR techniques so far developed, including strip biopsy, EMR using a cap, Makuuchi tube method, etc. The selection of EMR method largely depends on personal experience and preference of an endoscopist. Here we present our EMR method along with its advantages and disadvantages. With the extension of indications, more cases of early esophageal cancer will be treated by EMR. Since only long‐term results tell what the ideal EMR method is, well‐designed comparative studies are necessary in the future to establish a standard method.
Endoscopic Mucosal Resection for Esophageal Cancer: EMR‐C Procedure
Authors: YOSHIDA TATSUYA , INOUE HARUHIRO , KUDO SHIN‐EI
Published: July 2003

As a therapeutic modality for superficial esophageal cancer, endoscopic mucosal resection (EMR) is now thoroughly established. We routinely perform EMR using a cap (EMR‐C) procedure as a developer. In EMR‐C procedure, injection of a large volume of saline and a certain prelooping along the inner rim of the cap are vital issues for safe and reliable resection. In multi‐sessions of EMR, submucosal injection of saline prior to every session is essential, in order to avoid perforation. In such a manner, total circumferential EMR can be performed safely and easily by the EMR‐C procedure.
Endoscopic Resection of Tumor of Papilla Vater: Our Experiences
Authors: YASUDA KENJIRO , UNO KOJI , TANAKA KIYOHITO , NAKAJIMA MASATSUGU
Published: July 2003

Endoscopic papillectomy has become common in recent years as a non‐invasive therapeutic procedure for tumor of the papilla of Vater, although the lesions, which indicate endoscopic papillectomy, are very rare. In order to decide the indication for this procedure, intraductal ultrasonography (IDUS) is useful to obtain the correct diagnosis. For the successful and safe procedure of papillectomy, it is important to use the cutting mode of generator. and it is sometimes useful to inject the saline into the submucosal layer of the duodenum and to place the plastic stent into the bilio‐pancreatic ducts.
Quantification of dissimilatory (bi)sulphite reductase gene expression in Desulfobacterium autotrophicum using real‐time RT‐PCR
Authors: Neretin Lev N. , Schippers Axel , Pernthaler Annelie , Hamann Knut , Amann Rudolf , Jørgensen Bo Barker
Published: July 2003

We developed a real‐time RT‐PCR method for the quantification of dissimilatory (bi)sulphite reductase (DSR) mRNA in Desulfobacterium autotrophicum cells. The amount of DSR mRNA was determined relative to the amount of 16S rRNA at different growth conditions during transition from exponential to stationary phase: sulphate respiration with lactate, thiosulphate respiration with lactate, sulphate respiration with H2 and pyruvate fermentation. The dsr gene was expressed constitutively, although DSR mRNA content per‐cell varied under different growth conditions. The maximum DSR mRNA per‐cell content was 2.0 to 4.1‐fold higher during sulphate or thiosulphate respiration than during pyruvate fermentation. After transfer of a pyruvate‐fermenting culture into sulphate‐rich medium, upregulation of the DSR mRNA content was observed. Irrespective of the mode of metabolism the per‐cell DSR mRNA content changed significantly during growth (up to 310‐fold from the early to the late exponential phase during respiration with thiosulphate). The maximum DSR mRNA per‐cell contents correlated with cell‐specific sulphate reduction rates for all experiments. Environmental applications for the quantification of DSR mRNA are discussed.
Interspecific evolution: microbial symbiosis, endosymbiosis and gene transfer
Authors: Hoffmeister Meike , Martin William
Published: July 2003

Microbial symbioses are interesting in their own right and also serve as exemplary models to help biologists to understand two important symbioses in the evolutionary past of eukaryotic cells: the origins of chloroplasts and mitochondria. Most, if not all, microbial symbioses have a chemical basis: compounds produced by one partner are useful for the other. But symbioses can also entail the transfer of genes from one partner to the other, which in some cases cements two cells into a bipartite, co‐evolving unit. Here, we discuss some microbial symbioses in which progress is being made in uncovering the nature of symbiotic interactions: anaerobic methane‐oxidizing consortia, marine worms that possess endosymbionts instead of a digestive tract, amino acid‐producing endosymbionts of aphids, prokaryotic endosymbionts living within a prokaryotic host within mealybugs, endosymbionts of an insect vector of human disease and a photosynthetic sea slug that steals chloroplasts from algae. In the case of chloroplasts and mitochondria, examples of recent and ancient gene transfer to the chromosomes of their host cell illustrate the process of genetic merger in the wake of organelle origins.
Influence of the growth substrate on ester‐linked phospho‐ and glycolipid fatty acids of PAH‐degrading Mycobacterium sp. LB501T
Authors: Wick Lukas Y. , Pelz Oliver , Bernasconi Stefano M. , Andersen Nils , Harms Hauke
Published: July 2003

The influences of poorly water‐soluble anthracene on ester‐linked phospholipid fatty acid (PLFA) and glycolipid fatty acid (GLFA) profiles of Mycobacterium sp. LB501T were studied. Bacteria were cultivated on either anthracene or glucose (one culture with successively amended small doses of this substrate and one with excess concentrations) to distinguish between influences of the chemical structure and the bioavailability of the growth substrate. Results revealed that GLFA and PLFA profiles of M. sp. LB501T depended on the availability and the structure of the carbon source. Fatty acid profiles obtained with anthracene differed from those obtained with excess glucose. They were interpreted as a specific adaptation to this poorly bioavailable polycyclic aromatic hydrocarbon (PAH). In contrast, profiles obtained with low glucose concentrations showed clear signs of starvation stress. Stable carbon isotopic ratios (δ13C) of GLFA and PLFA of M. sp. LB501T were analysed to characterize the 13C‐fractionation during the biosynthesis of individual fatty acids and to evaluate their value as markers for substrate usage. Although the δ13C values of PLFA and GLFA showed differential isotope fractionation during anthracene‐ and glucose‐degradation, they were sufficiently distinct to be used as signatures of bacterial substrate usage.
Analysis of natural populations of Prochlorococcus spp. in the northern Red Sea using phycoerythrin gene sequences
Authors: Steglich Claudia , Post Anton F. , Hess Wolfgang R.
Published: July 2003

Marine cyanobacteria of the genus Prochlorococcus belong to one of two ecotypes that are specifically adapted to either low light (LL) or high light (HL) conditions. Previous analyses of the differences in pigmentation and gene complement revealed that LL‐adapted ecotypes carry a gene cluster to produce a functional phycoerythrin, whereas in the fully sequenced genome of the HL‐adapted strain MED4, only a single and free‐standing cpeB gene occurs. This gene encodes a derived form of β‐phycoerythrin, the function of which has remained enigmatic so far. Here, an analysis of HL‐adapted Prochlorococcus strains from different ocean provinces revealed the presence of a cpeB gene highly similar to that of MED4. To investigate whether the presence of particular phycoerythrin genes is a common characteristic of the LL‐ and HL‐adapted ecotypes, primer sets targeting specific motifs in LL‐cpeB and HL‐cpeB were designed for polymerase chain reaction (PCR) analysis of Red Sea phytoplankton. A major PCR product for Prochlorococcus HL‐cpeB was obtained from samples taken at 5–70 m depth and for LL‐cpeB from 70–125 m. The high sensitivity of this approach allowed the detection of HL‐cpeB down to 100 m and LL‐cpeB as deep as 175 m. DNA sequence and phylogenetic analysis of 70 individual clones for HL‐cpeB and of 68 clones for LL‐cpeB revealed a monophyletic origin for the HL and LL sequences respectively. This study shows that cpeB sequences are suitable as very sensitive molecular markers for the study of natural populations of Prochlorococcus. The low sequence divergence of HL‐cpeB among Prochlorococcus strains, which have been isolated from the Mediterranean Sea, the Arabian Sea and the Southern Pacific Ocean as well as in populations from the Red Sea, suggests the HL‐cpeB gene to be conserved and its product to be functional in Prochlorococcus.
Effects of temperature and fertilizer on activity and community structure of soil ammonia oxidizers
Authors: Avrahami Sharon , Liesack Werner , Conrad Ralf
Published: July 2003

We investigated the effect of temperature on the activity of soil ammonia oxidizers caused by changes in the availability of ammonium and in the microbial community structure. Both short (5 days) and long (6.5, 16 and 20 weeks) incubation of an agricultural soil resulted in a decrease in ammonium concentration that was more pronounced at temperatures between 10 and 25°C than at either 4°C or 30–37°C. Consistently, potential nitrification was higher between 10 and 25°C than at either 4°C or 37°C. However, as long as ammonium was not limiting, release rates of N2O increased monotonously between 4 and 37°C after short‐term temperature adaptation, with nitrification accounting for about 35–50% of the N2O production between 4 and 25°C. In order to see whether temperature may also affect the community structure of ammonia oxidizers, we studied moist soil during long incubation at low and high concentrations of commercial fertilizer. The soil was also incubated in buffered (pH 7) slurry amended with urea. Communities of ammonia oxidizers were assayed by denaturant gradient gel electrophoresis (DGGE) of the amoA gene coding for the α subunit of ammonia monooxygenase. We found that a polymerase chain reaction (PCR) system using a non‐degenerated reverse primer (amoAR1) gave the best results. Community shifts occurred in all soil treatments after 16 weeks of incubation. The community shifts were obviously influenced by the different fertilizer treatments, indicating that ammonium was a selective factor for different ammonia oxidizer populations. Temperature was also a selective factor, in particular as community shifts were also observed in the soil slurries, in which ammonium concentrations and pH were better controlled. Cloning and sequencing of selected DGGE bands indicated that amoA sequences belonging to Nitrosospira cluster 1 were dominant at low temperatures (4–10°C), but were absent after long incubation at low fertilizer treatment. Sequences of Nitrosospira cluster 9 could only be detected at low ammonium concentrations, whereas those of Nitrosospira cluster 3 were found at most ammonium concentrations and temperatures, although individual clones of this cluster exhibited trends with temperature. Obviously, ammonia oxidizers are able to adapt to soil conditions by changes in the community structure if sufficient time (several weeks) is available.
Studies on pathogenic Vibrio parahaemolyticus during a warm weather season in the Seto Inland Sea, Japan
Authors: Alam M. Jahangir , Miyoshi Shin‐Ichi , Shinoda Sumio
Published: July 2003

Vibrio parahaemolyticus is a potentially pathogenic bacterium, occurring naturally in estuarine and marine environments throughout the world. The incidence of this organism in an aquatic environment depends upon many ecofactors. Sea water and organic material were collected during the warm weather season from a coast of the Seto Inland Sea, Japan, and analysed to determine V. parahaemolyticus densities and the occurrence of pathogenic strains, defined as those possessing tdh and/or trh genes by polymerase chain reaction (PCR), using isolated DNA from enrichment culture of the samples. About 99% of samples were positive for V. parahaemolyticus with densities of 3 to> 1400 cells per 100 ml of water or 10 g of organic samples by the most‐probable‐number (MPN)‐PCR technique, but only 76.6% were positive by the conventional MPN culture technique, with densities ranging from 3 to> 1400 cells per 100 ml of water or 10 g of organics. Furthermore, the tdh and trh genes were positive in 41.5% and 8.5% of samples, respectively, by the MPN‐PCR technique. No tdh and trh gene‐positive strains were isolated by the conventional MPN culture procedure. The difference in detection between the MPN culture and the MPN‐PCR techniques appeared to be significant and may be attributed to different detection sensitivities and other factors.
Impact of antibiotics on conjugational resistance gene transfer in Staphylococcus aureus in sewage
Authors: Ohlsen Knut , Ternes Thomas , Werner Guido , Wallner Ursula , Löffler Dirk , Ziebuhr Wilma , Witte Wolfgang , Hacker Jörg
Published: July 2003

The growing rate of microbial pathogens becoming resistant to standard antibiotics is an important threat to public health. In order to assess the role of antibiotics in the environment on the spread of resistance factors, the impact of subinhibitory concentrations of antibiotics in sewage on gene transfer was investigated using conjugative gentamicin resistance (aacA–aphD) plasmids of Staphylococcus aureus. Furthermore, the concentration of antibiotics in hospital sewage was measured by high‐performance liquid chromatography (HPLC)–electrospray tandem mass spectrometry. Several antibiotics were found to be present in sewage, e.g. ciprofloxacin up to 0.051 mg l−1 and erythromycin up to 0.027 mg l−1. Resistance plasmid transfer occurred both on solidified (dewatered) sewage and in liquid sewage in a bioreactor with a frequency of 1.1 × 10−5−5.0 × 10−8. However, low‐level concentrations of antibiotics measured in sewage are below concentrations that can increase plasmid transfer frequencies of gentamicin resistance plasmids of staphylococci.