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Spirochaetes in small bowel
gel of the PCR amplification product was then Southern blot transferred and hybridised for the published 32P end-labelled 25 basepair internal oligonucleotide probe. Our results revealed that all 4 AIDS-KS specimens (3 skin lesions and 1 lymph node) were positive for this herpesviruslike signal. Importantly, 2 of the 3 non-AIDS KS biopsies (1 skin lesion; 1 lymph node KS lesion) were also positive for this herpesvirus-like DNA sequence. The remaining 5 AIDS lymph nodes and the 32 non-AIDS lymphoid lesions were all negative for this DNA sequence despite prolonged exposure of the autoradiographed membrane. Our preliminary studies from a southeastern Asian region are therefore in support of the finding that the herpesviruslike DNA sequences represent a distinct viral agent closely associated with the development of KS either in AIDS patients or in non-AIDS patients. Sequence analysis and comparison with that in published data from western countries will be continued. agarose
*Ih-Jen
Su, Yeong-Shian Hsu, Yun-Chuun Chang, I-Wen Wang
Department of
Pathology, National Cheng-Kung University Hospital and College of Medicine, Taiwan; Department of Pathology, Tzu-Chi General Hospital, Hua-Lien; and *Department of Pathology, National Taiwan University Hospital, Taipei, Taiwan 1 Drew WL, Conant MA, Miner RC, et al. Cytomegalovirus and Kaposi’s sarcoma in young homosexual men. Lancet 1982; ii: 1255-27. 2 Vogel J, Hinrichs SH, Reynolds RK, Luciw PA, Jay G. The HIV tat gene induces dermal lesions resembling Kaposi’s sarcoma in transgenic mice. Nature 1988; 335: 606-11. 3 Bovenzi P, Mirandola P, Secchiero P, et al. Human herpesvirus 6 (variant A) in Kaposi’s sarcoma. Lancet 1993; 341: 1288-89. 4 Chang Y, Cesarman E, Pessin MS, et al. Identification of herpesviruslike DNA sequences in AIDS-associated Kaposi’s sarcoma. Science 5
1994; 266: 1865-69. Su IJ, Hsieh HC, Lin KH, et al. Aggressive peripheral T cell lymphomas containing Epstein-Barr viral DNA: a clinicopathologic and molecular analysis. Blood 1991; 77: 799-808.
Figure: Spirochaetes in small bowel on mucosa of small Intestine taken from about 60 cm distal to the pyloric sphincter, from patient with RA. Reference bar is 1 um.
A=Slngle spirochaetal organism, lying freely
observed collection of spiral organisms, lying freely on surface of small bowel mucosa. They were never seen to be associated with mucosal damage, and none was seen to invade mucosa. Reference bar is 5 um.
B=Commonly
these organisms. Our findings contradict this and seem extend spirochaetes’ range of habitat. We propose that the human small bowel, like the large bowel, is a common site where spirochaetes may be found. Although a number of our subjects had forms of arthritis and we are aware that at least one form of arthritis is associated with a spirochaetal organism, we do not presently propose that these small bowel spirochaetes are associated with the pathogenesis of RA. The organisms remain to be fully identified, and their possible pathological significance has yet to be explained.
by to
Ursula Potter, *A J Collins, Lidia J Notarianni, A John Morris, A W Smith Electron Optics Unit, University of Bath, Bath, *School of Pharmacy and Pharmacology, University of Bath, Bath BA2 7AY, UK; and Department of Gastroenterology, Royal Infirmary, Glasgow
1
Spirochaetes in
small bowel
of the small bowel by enteroscopy to record damage caused by nonsteroidal anti-inflammatory drugs. Incidentally, we have found that mucosal biopsy samples taken on these occasions, when examined by scanning electron microscope, have shown the presence of spirochaetal organisms. 14 patients with rheumatoid arthritis (RA), 1 patient with ankylosing spondylitis, and 5 patients with a variety of gastrointestinal complaints were studied. Biopsy specimens were taken via an enteroscope from, on average, 60 cm distal to. the pyloric sphincter. The samples were immediately fixed in 2% glutaraldelyde/0-05 mol/L phosphate buffer, pH 7-4, for 24 h, and then post-fixed with osmium tetroxide in 0-11 mol/L phosphate/0-15 mol/L sucrose buffer followed by OTOTO method of fixation. 1,2 The specimens were examined with a JEOL JSM-T330 scanning electron SiR-We have been
examining
the
mucosa
microscope. Of the patients studied, 7 of the patients with RA (50%), the patient with ankylosing spondylitis, and 4 of the 5 other subjects showed the presence of spirochaetal-like organisms lying on the surface of the intestinal mucosa. They varied in length from 10-30 (im. Some of these organisms were found singly, and others in groups (figure). It is not possible to know to what extent the process of tissue fixation removed any of these organisms from the mucosal surface, but it seems clear that those remaining were found so commonly in random biopsy samples taken from a variety of subjects that spirochaetes must be organisms that commonly inhabit the small bowel. Although spirochaetes are frequently found in the mouth and large bowel’ the small bowel has not been reported to be colonised
2
3
4
Seligman AM, Wasserkrug HL, Hanker JS. A new staining method (OTO) for enhancing contrast of lipid-containing membranes and droplets in osmium tetroxide fixed tissue with osmiophilic thio carbiohydrazide (TCH). J Cell Biol 1966; 30: 424-32. Malick LE, Wilson RB. Evaluation of a modified technique for SEM examination of vertebrate specimens without evaporated metal layers. Scanning Electron Microsc 1975; 1: 259-66. Willen R, Carlen B, Cronstedt J, Willen H. Intestinal spirochaetosis of the colon diagnosed with colonoscopy and multiple biopsies. Endoscopy 1985; 17: 86-88. PJ, Nakala MM, Reinhart JF, George WL. Spirochete-like organisms in the human gastrointestinal tract. Rev Infect Dis 1989;
Ruane
11:
184-95.
Genetic abnormalities
during transition from Helicobacter-pylori-associated gastritis to low-grade MALToma
SiR-On the assumption that Helicobacter pylori infection is critical in the pathogenesis of low-grade B-cell gastric lymphoma of the MALT type, Calvert and colleagues (Jan 7, p 26) investigate allele imbalance at six tumoursuppressor gene loci as an indicator of progression from H pylori-associated gastritis to low-grade B-cell lymphoma of the MALT type. Although this is a major contribution to the understanding of the molecular pathogenesis of gastric lymphoma, we believe that additional factors should be taken into consideration. The assumed pathogenic role of H pylori in initiating and promoting the disease is not evident, for several reasons. First, only one study, in 3 patients, shows T-cell dependent B-cell
proliferation in response to H pylori.’ Second, the reported remission of primary gastric low-grade B-cell lymphoma of the MALT type in 5 of 6 patients by eradication of H pylori is hampered by absence of clinical 723
*Ih-Jen
Su, Yeong-Shian Hsu, Yun-Chuun Chang, I-Wen Wang
Department of
Pathology, National Cheng-Kung University Hospital and College of Medicine, Taiwan; Department of Pathology, Tzu-Chi General Hospital, Hua-Lien; and *Department of Pathology, National Taiwan University Hospital, Taipei, Taiwan 1 Drew WL, Conant MA, Miner RC, et al. Cytomegalovirus and Kaposi’s sarcoma in young homosexual men. Lancet 1982; ii: 1255-27. 2 Vogel J, Hinrichs SH, Reynolds RK, Luciw PA, Jay G. The HIV tat gene induces dermal lesions resembling Kaposi’s sarcoma in transgenic mice. Nature 1988; 335: 606-11. 3 Bovenzi P, Mirandola P, Secchiero P, et al. Human herpesvirus 6 (variant A) in Kaposi’s sarcoma. Lancet 1993; 341: 1288-89. 4 Chang Y, Cesarman E, Pessin MS, et al. Identification of herpesviruslike DNA sequences in AIDS-associated Kaposi’s sarcoma. Science 5
1994; 266: 1865-69. Su IJ, Hsieh HC, Lin KH, et al. Aggressive peripheral T cell lymphomas containing Epstein-Barr viral DNA: a clinicopathologic and molecular analysis. Blood 1991; 77: 799-808.
Figure: Spirochaetes in small bowel on mucosa of small Intestine taken from about 60 cm distal to the pyloric sphincter, from patient with RA. Reference bar is 1 um.
A=Slngle spirochaetal organism, lying freely
observed collection of spiral organisms, lying freely on surface of small bowel mucosa. They were never seen to be associated with mucosal damage, and none was seen to invade mucosa. Reference bar is 5 um.
B=Commonly
these organisms. Our findings contradict this and seem extend spirochaetes’ range of habitat. We propose that the human small bowel, like the large bowel, is a common site where spirochaetes may be found. Although a number of our subjects had forms of arthritis and we are aware that at least one form of arthritis is associated with a spirochaetal organism, we do not presently propose that these small bowel spirochaetes are associated with the pathogenesis of RA. The organisms remain to be fully identified, and their possible pathological significance has yet to be explained.
by to
Ursula Potter, *A J Collins, Lidia J Notarianni, A John Morris, A W Smith Electron Optics Unit, University of Bath, Bath, *School of Pharmacy and Pharmacology, University of Bath, Bath BA2 7AY, UK; and Department of Gastroenterology, Royal Infirmary, Glasgow
1
Spirochaetes in
small bowel
of the small bowel by enteroscopy to record damage caused by nonsteroidal anti-inflammatory drugs. Incidentally, we have found that mucosal biopsy samples taken on these occasions, when examined by scanning electron microscope, have shown the presence of spirochaetal organisms. 14 patients with rheumatoid arthritis (RA), 1 patient with ankylosing spondylitis, and 5 patients with a variety of gastrointestinal complaints were studied. Biopsy specimens were taken via an enteroscope from, on average, 60 cm distal to. the pyloric sphincter. The samples were immediately fixed in 2% glutaraldelyde/0-05 mol/L phosphate buffer, pH 7-4, for 24 h, and then post-fixed with osmium tetroxide in 0-11 mol/L phosphate/0-15 mol/L sucrose buffer followed by OTOTO method of fixation. 1,2 The specimens were examined with a JEOL JSM-T330 scanning electron SiR-We have been
examining
the
mucosa
microscope. Of the patients studied, 7 of the patients with RA (50%), the patient with ankylosing spondylitis, and 4 of the 5 other subjects showed the presence of spirochaetal-like organisms lying on the surface of the intestinal mucosa. They varied in length from 10-30 (im. Some of these organisms were found singly, and others in groups (figure). It is not possible to know to what extent the process of tissue fixation removed any of these organisms from the mucosal surface, but it seems clear that those remaining were found so commonly in random biopsy samples taken from a variety of subjects that spirochaetes must be organisms that commonly inhabit the small bowel. Although spirochaetes are frequently found in the mouth and large bowel’ the small bowel has not been reported to be colonised
2
3
4
Seligman AM, Wasserkrug HL, Hanker JS. A new staining method (OTO) for enhancing contrast of lipid-containing membranes and droplets in osmium tetroxide fixed tissue with osmiophilic thio carbiohydrazide (TCH). J Cell Biol 1966; 30: 424-32. Malick LE, Wilson RB. Evaluation of a modified technique for SEM examination of vertebrate specimens without evaporated metal layers. Scanning Electron Microsc 1975; 1: 259-66. Willen R, Carlen B, Cronstedt J, Willen H. Intestinal spirochaetosis of the colon diagnosed with colonoscopy and multiple biopsies. Endoscopy 1985; 17: 86-88. PJ, Nakala MM, Reinhart JF, George WL. Spirochete-like organisms in the human gastrointestinal tract. Rev Infect Dis 1989;
Ruane
11:
184-95.
Genetic abnormalities
during transition from Helicobacter-pylori-associated gastritis to low-grade MALToma
SiR-On the assumption that Helicobacter pylori infection is critical in the pathogenesis of low-grade B-cell gastric lymphoma of the MALT type, Calvert and colleagues (Jan 7, p 26) investigate allele imbalance at six tumoursuppressor gene loci as an indicator of progression from H pylori-associated gastritis to low-grade B-cell lymphoma of the MALT type. Although this is a major contribution to the understanding of the molecular pathogenesis of gastric lymphoma, we believe that additional factors should be taken into consideration. The assumed pathogenic role of H pylori in initiating and promoting the disease is not evident, for several reasons. First, only one study, in 3 patients, shows T-cell dependent B-cell
proliferation in response to H pylori.’ Second, the reported remission of primary gastric low-grade B-cell lymphoma of the MALT type in 5 of 6 patients by eradication of H pylori is hampered by absence of clinical 723