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Organ culture of mouse vibrissal follicles. 1. Effect of anticancer agent on DNA synthesis
J55
556
PATHOGENESIS
OF HIV-1
IN PERIPHERAL
BLOOD
MONONUCLEAR
CELLS
Time cowse of neutrophil infiltration staphylococcal impetigo in mice
MICHIYO SHIBATAl).SHUHEI SHIMAO1),TAKASHI KURIfilJRAz)~ 1)Department of Dermatology, Faculty of Nedicine, Tottori Univ. StDepartment of Pathology. Research Institute for Microbial Diseases, Osaka Univ.
in experimental
of human immunodeficiency virus type-l(HIV-1) in human peripheral blood mononuclear a specified healthy donor as the highly permissive cell for the organism. After infection with HIV strains ohtaind from carriers with various clinical states. the viability of PBMCas an indicator of pathogenicity and. clinical state were found be correlated. A tendency for most strains highly pathogenic in vitro to have been isolated from symptomatic cases and for most of the less pathogenic strains to have been isolated from asymptomatic carriers was observed. Results of an additional experiment indicated that viruses less pathogenic in PBMCfrom a specified individual sometimes showed cytotoxic effects in PBMCfrom other individuals. This finding suggests that cellular factors influence viral pathogenicity.
YOSHIKO ABE, HISANORI AKIYAMA, JIRO ARATA Department of Dermatology, Okayama Univ. Medical School, Okayama The distribution of s.aureus in the cornea1 layer and epidermis and its time cowse were observed in an experimental impetigo model. S.aureus isolated from a human impetigo was applied on the slightly abraded back skin of female ddy micecddy-m) and C57BL/KsJdb/db mice(db/#b-m), and the inoculated areas were occluded under Uniflex Development of intraepidermal vesicles with many neutrophils was compared in db/db-m and ddy-m. Inoculation size required was smaller in db/db-m(7 week, blood sugar 361.3 mg/dl) than in ddy-m.Neutrophil infiltration after the inoculation was less marked-in db/db-m than in ddy/m. On rechallenge with s.aureus bv inoculation size 2.5~10~1 /0.05ml, db/db-m(ll week, blood sugar 481.4mgldlj "eutrophi1 infiltration was absent. In contrast neutrophils infiltration was more prominent than on first challenge with s.aureus i" ddy-m(ll week, blood sugar 110.1 mg/dl).
557
558
Pathogenesis was evaluated
cells(PBF1C) from
A RAPID PCR AND
DIAGNOSIS OF NONRADIOACTIVE
TSUTSUGAMUSHI DETECTION
DISEASE SYSTEM
.
USING
NON-INVASIVE MEASUREMENTS DENSITY CHANGES IN NORMAL AN ULTRASONIC SCANNER
YASUYUKI SUGITA, TETSUO SASAKI, TETSUO NAGATANI, NORIHISA ISHII AND HIROSHI NAKAJIMA Department of Dermatology, Yokohama City Univ. School of Medicine, Yokohama A polymerase chain reaction (PCR) usinq patients' blood 1s demonstrated for the specific detection of Rickettsia tsutsuqamushi, the causative aqent of scrub typhus (tsutsugamushi disease). TWO-
MASAHIRO NISHIMURA AND TAKUO Department of Dermatology, MedIcal School, Nagoya
OF AND
SKIN THICKNESS ABNORMAL SKIN
TSUJI Nagoya
and immunoqenic protein found to be homologous to HSP 60.- For a-practical diagnosis, amplified DNA was detected with a nonradioactive DNA probe. This study is enable to make a rapid diagnosis of tsutsugamushi disease during acute phase of disease.
J59
560
AN APPLICATION FOR A COLOR IMAGE ANALYZER FOR GOLD LABELLIG IN IMMUNOELECTRTON MICROSCOPY TO ACRIEVE ENHANCED DEMYONSTRATION OF SHALL GOLD PROBES.
ORGAN CULTURE OF MOUSE VIBRISSAL FOLLICLES. ANTICANCER AGENT ON DNA SYNTHESIS
School
Unlverslty
Using an ultrasonic scanner (Dermascan C) we measured the thxkness and density of the skin of several healthy young a"d old adults, PSS, and scars. The dermal thickness was significantly thinner 1" the old than 1" I" addition, a the young. band-like low dense areas were see" in the upper dermis of the sun-exposed skin (forearm and fk&) of the old. The chances in thickness and densitv of the dermis were also _ seen in PSS and sea; tissues. From these findings, we found that the ultrasonic scanner is a useful tool for measuring the skin thickness and some abnormal skin area.
Sets of primers were selected on the basis of the nucleotide sequence of a gene encoding the 58 kD protein, one of the most abundant
ffIROSR1 SRIMIZU AND TAKE.,1 NISHIRAWA Department of Dermatology, Keio University Medicine, Tokyo.
City
AND USING
1. EFFECT
OF
TOSHIMASA .,INDO,RY"S"KE IMAI, KENJI TAKAMORI and HIDEOKI OGAWA Department of Dermatology, Juntendo Univ. School of Medicine, Tokyo
of
We established a method for organ culture of mouse vibrissal follicles. Vibrissae were harvested from Fl hybrid mice (C57BL6XC3H) at g-day of age. Culture conditions of 5% COz 95% O2 at 31'C were found to be suitable for mouse vibrissal follicle, and utilyzing these conditions we successfully organ cultured mouse vibrissal follicle in serum-free media. The vibrissal hair bulb and germinative cells maintained their normal morophologies for 96 hr culture. Autoradiographys of "H-thymidine labeled follicles showed localization in the germinative cells below Auber's critical line. Vibrissal hair bulb DNA synthesis increased time dependently for 72 hr after culture initiation. We investigated the effect of some anticancer agents on the DNA synthesis of hair germinative cells using this system. Such an organ culture method in serum free media may be useful for studies on hair growth.
In iamunoelectron microscopy (IEM). immunogold has become one of the most valuable probes for labelling. The smaller the size of the the gold probes used in IEM. the denser inmunolabelling can be obtained. Therefore 5nm inmunogold. the smallest gold probes commerciallr available that are detectable under transmission EM. have been widely used for various studies. 5nm gold too small to be identified at loa probes are, however. magnification in transmission EM. We therefore spplied a color image analyzer (Olympus SP500) to detect individual gold particles more easily in IEM. Each individual 5nm gold particle that was enhanced in size and color is large snd evident enough for evaluating imnunolabelling even at low magnification. We feel that this new application of a color image analyzer in IEM can provide a better resolution in IEH.
240
556
PATHOGENESIS
OF HIV-1
IN PERIPHERAL
BLOOD
MONONUCLEAR
CELLS
Time cowse of neutrophil infiltration staphylococcal impetigo in mice
MICHIYO SHIBATAl).SHUHEI SHIMAO1),TAKASHI KURIfilJRAz)~ 1)Department of Dermatology, Faculty of Nedicine, Tottori Univ. StDepartment of Pathology. Research Institute for Microbial Diseases, Osaka Univ.
in experimental
of human immunodeficiency virus type-l(HIV-1) in human peripheral blood mononuclear a specified healthy donor as the highly permissive cell for the organism. After infection with HIV strains ohtaind from carriers with various clinical states. the viability of PBMCas an indicator of pathogenicity and. clinical state were found be correlated. A tendency for most strains highly pathogenic in vitro to have been isolated from symptomatic cases and for most of the less pathogenic strains to have been isolated from asymptomatic carriers was observed. Results of an additional experiment indicated that viruses less pathogenic in PBMCfrom a specified individual sometimes showed cytotoxic effects in PBMCfrom other individuals. This finding suggests that cellular factors influence viral pathogenicity.
YOSHIKO ABE, HISANORI AKIYAMA, JIRO ARATA Department of Dermatology, Okayama Univ. Medical School, Okayama The distribution of s.aureus in the cornea1 layer and epidermis and its time cowse were observed in an experimental impetigo model. S.aureus isolated from a human impetigo was applied on the slightly abraded back skin of female ddy micecddy-m) and C57BL/KsJdb/db mice(db/#b-m), and the inoculated areas were occluded under Uniflex Development of intraepidermal vesicles with many neutrophils was compared in db/db-m and ddy-m. Inoculation size required was smaller in db/db-m(7 week, blood sugar 361.3 mg/dl) than in ddy-m.Neutrophil infiltration after the inoculation was less marked-in db/db-m than in ddy/m. On rechallenge with s.aureus bv inoculation size 2.5~10~1 /0.05ml, db/db-m(ll week, blood sugar 481.4mgldlj "eutrophi1 infiltration was absent. In contrast neutrophils infiltration was more prominent than on first challenge with s.aureus i" ddy-m(ll week, blood sugar 110.1 mg/dl).
557
558
Pathogenesis was evaluated
cells(PBF1C) from
A RAPID PCR AND
DIAGNOSIS OF NONRADIOACTIVE
TSUTSUGAMUSHI DETECTION
DISEASE SYSTEM
.
USING
NON-INVASIVE MEASUREMENTS DENSITY CHANGES IN NORMAL AN ULTRASONIC SCANNER
YASUYUKI SUGITA, TETSUO SASAKI, TETSUO NAGATANI, NORIHISA ISHII AND HIROSHI NAKAJIMA Department of Dermatology, Yokohama City Univ. School of Medicine, Yokohama A polymerase chain reaction (PCR) usinq patients' blood 1s demonstrated for the specific detection of Rickettsia tsutsuqamushi, the causative aqent of scrub typhus (tsutsugamushi disease). TWO-
MASAHIRO NISHIMURA AND TAKUO Department of Dermatology, MedIcal School, Nagoya
OF AND
SKIN THICKNESS ABNORMAL SKIN
TSUJI Nagoya
and immunoqenic protein found to be homologous to HSP 60.- For a-practical diagnosis, amplified DNA was detected with a nonradioactive DNA probe. This study is enable to make a rapid diagnosis of tsutsugamushi disease during acute phase of disease.
J59
560
AN APPLICATION FOR A COLOR IMAGE ANALYZER FOR GOLD LABELLIG IN IMMUNOELECTRTON MICROSCOPY TO ACRIEVE ENHANCED DEMYONSTRATION OF SHALL GOLD PROBES.
ORGAN CULTURE OF MOUSE VIBRISSAL FOLLICLES. ANTICANCER AGENT ON DNA SYNTHESIS
School
Unlverslty
Using an ultrasonic scanner (Dermascan C) we measured the thxkness and density of the skin of several healthy young a"d old adults, PSS, and scars. The dermal thickness was significantly thinner 1" the old than 1" I" addition, a the young. band-like low dense areas were see" in the upper dermis of the sun-exposed skin (forearm and fk&) of the old. The chances in thickness and densitv of the dermis were also _ seen in PSS and sea; tissues. From these findings, we found that the ultrasonic scanner is a useful tool for measuring the skin thickness and some abnormal skin area.
Sets of primers were selected on the basis of the nucleotide sequence of a gene encoding the 58 kD protein, one of the most abundant
ffIROSR1 SRIMIZU AND TAKE.,1 NISHIRAWA Department of Dermatology, Keio University Medicine, Tokyo.
City
AND USING
1. EFFECT
OF
TOSHIMASA .,INDO,RY"S"KE IMAI, KENJI TAKAMORI and HIDEOKI OGAWA Department of Dermatology, Juntendo Univ. School of Medicine, Tokyo
of
We established a method for organ culture of mouse vibrissal follicles. Vibrissae were harvested from Fl hybrid mice (C57BL6XC3H) at g-day of age. Culture conditions of 5% COz 95% O2 at 31'C were found to be suitable for mouse vibrissal follicle, and utilyzing these conditions we successfully organ cultured mouse vibrissal follicle in serum-free media. The vibrissal hair bulb and germinative cells maintained their normal morophologies for 96 hr culture. Autoradiographys of "H-thymidine labeled follicles showed localization in the germinative cells below Auber's critical line. Vibrissal hair bulb DNA synthesis increased time dependently for 72 hr after culture initiation. We investigated the effect of some anticancer agents on the DNA synthesis of hair germinative cells using this system. Such an organ culture method in serum free media may be useful for studies on hair growth.
In iamunoelectron microscopy (IEM). immunogold has become one of the most valuable probes for labelling. The smaller the size of the the gold probes used in IEM. the denser inmunolabelling can be obtained. Therefore 5nm inmunogold. the smallest gold probes commerciallr available that are detectable under transmission EM. have been widely used for various studies. 5nm gold too small to be identified at loa probes are, however. magnification in transmission EM. We therefore spplied a color image analyzer (Olympus SP500) to detect individual gold particles more easily in IEM. Each individual 5nm gold particle that was enhanced in size and color is large snd evident enough for evaluating imnunolabelling even at low magnification. We feel that this new application of a color image analyzer in IEM can provide a better resolution in IEH.
240