Wound healing in diabetic ulcers remains a significant clinical challenge,primarily due to bacterial infection and impaired angiogenesis.Periplaneta americana extract(PAE)has been widely used to treat diabetic wounds,...Wound healing in diabetic ulcers remains a significant clinical challenge,primarily due to bacterial infection and impaired angiogenesis.Periplaneta americana extract(PAE)has been widely used to treat diabetic wounds,yet its underlying mechanisms are not fully understood.This study aimed to elucidate these mechanisms by analyzing long non-coding RNA(lncRNA)expressions in the wound tissues from diabetic anal fistula patients treated with or without PAE,using high-throughput sequencing.Peripheral blood monocytes from patients were differentiated into M0 macrophages with human macrophage colony-stimulating factor(hMCSF)and subsequently polarized into M1 macrophages with lipopolysaccharide.The results indicated that LINC01133 and SLAMF9 were downregulated in wound tissues of patients treated with PAE.Furthermore,PAE suppressed M1 macrophage polarization and enhanced human umbilical vein endothelial cell(HUVEC)proliferation,migration,and angiogenesis.These effects were diminished when LINC01133 or SLAMF9 were overexpressed.Mechanistically,LINC01133 was shown to upregulate SLAMF9 through interaction with ELAVL1.Overexpression of SLAMF9 reversed the effects of LINC01133 silencing on macrophage polarization and HUVEC functions.In conclusion,PAE facilitates the healing of infected diabetic ulcers by downregulating the LINC01133/SLAMF9 pathway.展开更多
Researches on pharmacological action and drug development of Periplane- ta americana have demonstrated that it has wide medicinal value, low side effects and rich resources, having good application prospects in medici...Researches on pharmacological action and drug development of Periplane- ta americana have demonstrated that it has wide medicinal value, low side effects and rich resources, having good application prospects in medicine field. This article reviewed the researches on pharmacological action and clinical application of Periplaneta americana in order to provide references for the further research and devel- opment of Periplaneta americana medicines.展开更多
OBJECTIVE： To investigate the effect of the periplaneta americana extract on the intestinal mucosal barrier and prognostic implications in patients with sepsis. METHODS： Sixty and six patients with sepsis were assig...OBJECTIVE： To investigate the effect of the periplaneta americana extract on the intestinal mucosal barrier and prognostic implications in patients with sepsis. METHODS： Sixty and six patients with sepsis were assigned randomly to treatment group （32 cases） and control group （32 cases）. The extractfrom periplaneta americana plus conventional medication for sepsis was administered to the treatment group, while the control group only received conventional treatment. The gastrointestinal function scores and acute physiology and chronic health evaluation II （APACHE 11 ） scores of all subjects were documented at baseline, at days 1, 3 and 7 after treatment respectively and their blood endotoxin was tested at the same time points as well. The incidence of death was recorded for both groups throughout the trial. RESULTS： At days 3 and 7 after treatment, gastrointestinal function score, APACHE II, and endotoxin level in treatment group wasbetter than that in con- trol group and the difference between them was significant （both P〈0~05）. Although the incidence of death in treatment group was less than that in control group, the difference between the two groups was not significant （P〉 0.05）. CONCLUSION： The extract of periplaneta ameri- cana had protective effect on intestinal mucosal barrier and could improve the condition and prog- nosis in patients with sepsis.展开更多
BACKGROUND:Periplaneta americana extract is recognized to have a positive effect on gastrointestinal mucosa.This study aimed to investigate the effects of periplaneta americana extract on immune function,nutrition sta...BACKGROUND:Periplaneta americana extract is recognized to have a positive effect on gastrointestinal mucosa.This study aimed to investigate the effects of periplaneta americana extract on immune function,nutrition status and gastrointestinal complications of early enteral nutrition patients with systemic inflammatory response syndrome(SIRS).METHODS:Patients with SIRS were randomly divided into two groups:treatment and control groups.All patients in the two groups received conventional therapy including enteral nutrition,but periplaneta americana extract,an additional Chinese medicine,was given to the patients in the treatment group.At the beginning of treatment(0 day)and 1,3,and 7 days after treatment,the levels of immunoglobulin(Ig A),total lymphocyte count(TLC),total protein(TP)and prealbumin(PA)were respectively tested in patients'venous blood.The incidences of bloating,diarrhea,aspiration pneumonia and high blood sugar at 7 days after treatment were recorded.The mortality of the patients in 28 days was recorded.RESULTS:At 3 and 7 days after treatment,the levels of Ig A and TLC in the treatment group were higher than those in the control group(P<0.05).At 7 days after treatment,the levels of TP and PA in the treatment group were higher than those in the control group(P<0.05).The incidences of bloating and diarrhea in the treatment group were lower than those in the control group,the differences were significant(P<0.05).The mortality of treatment group was lower than that of the control group(P>0.05).CONCLUSION:Periplaneta americana extract could reduce gastrointestinal complications and improve immune function and nutritional status in patients with systemic inflammatory response syndrome.展开更多
Objective Acute lung injury(ALI)is an acute clinical syndrome characterized by uncontrolled inflammation response,which causes high mortality and poor prognosis.The present study determined the protective effect and u...Objective Acute lung injury(ALI)is an acute clinical syndrome characterized by uncontrolled inflammation response,which causes high mortality and poor prognosis.The present study determined the protective effect and underlying mechanism of Periplaneta americana extract(PAE)against lipopolysaccharide(LPS)-induced ALI.Methods The viability of MH-S cells was measured by MTT.ALI was induced in BALB/c mice by intranasal administration of LPS(5 mg/kg),and the pathological changes,oxidative stress,myeloperoxidase activity,lactate dehydrogenase activity,inflammatory cytokine expression,edema formation,and signal pathway activation in lung tissues and bronchoalveolar lavage fluid(BALF)were examined by H&E staining,MDA,SOD and CAT assays,MPO assay,ELISA,wet/dry analysis,immunofluorescence staining and Western blotting,respectively.Results The results revealed that PAE obviously inhibited the release of proinflammatory TNF-α,IL-6 and IL-1βby suppressing the activation of MAPK/Akt/NF-κB signaling pathways in LPS-treated MH-S cells.Furthermore,PAE suppressed the neutrophil infiltration,permeability increase,pathological changes,cellular damage and death,pro-inflammatory cytokines expression,and oxidative stress upregulation,which was associated with its blockage of the MAPK/Akt/NF-κB pathway in lung tissues of ALI mice.Conclusion PAE may serve as a potential agent for ALI treatment due to its anti-inflammatory and anti-oxidative properties,which correlate to the blockage of the MAPK/NF-κB and AKT signaling pathways.展开更多
[Objective] The aim of this study was to investigate the decolorization ef- fects of activated carbon and macroporous resin on Periplaneta americana L. skimmed cream and compare the advantages and disadvantages of the...[Objective] The aim of this study was to investigate the decolorization ef- fects of activated carbon and macroporous resin on Periplaneta americana L. skimmed cream and compare the advantages and disadvantages of the two decolorization technologies. [Method] Periplaneta americana L. skimmed cream was decolored with activated carbon and macroporous resin, and freeze-dried to collect solid decolorization products. By investigating the yield, decolorization rate, protein retention rate and decolorization operation process, the advantages and disadvantages of the two decolorization technologies were compared. [Result] Both activated carbon and macroporous resin can be used for decolorization. To be specific, macroporous resin-decolorization is superior in the yield and protein retention rate, while activated carbon-decolorization is superior in decolorization rate and decolorization operation process. [Conclusion] Macroporous resin-decolorization can be used if protein is the main ingredient required in the experiment, while activated carbon-decolorization can be used if protein is not the main ingredient required.展开更多
[ Objective] This study aimed to explore a Tricine-SDS-PAGE method for separation and purification of Periplaneta Americana small peptides. [ Method] By comparing the separation effects of three types of gels with dif...[ Objective] This study aimed to explore a Tricine-SDS-PAGE method for separation and purification of Periplaneta Americana small peptides. [ Method] By comparing the separation effects of three types of gels with different ratios on P. Americana small peptides, an appropriate Tricine-SDS-PAGE method for separation of small peptides was established. [ Result] Standard low molecular weight Marker was preliminarily separated to a certain extent with polyacrylamide gel electraphoresis, while the sample mixture was not separated due to the different levels of aggregation in separating gel, spacer gel and stacking gel. [Condusion] More appropriate polyacrylamide gel electrephoresis separation method or other better separation methods were to be further explored for separating peptide mixture with smaller molecular weight.展开更多
[Objective] The paper was to establish ultrafiltration membrane technology for separation of antitumor active plypeptides from Periplaneta Americana. [Method] Hollow fiber ultrafiltration membrane（3 kDa）was used to ...[Objective] The paper was to establish ultrafiltration membrane technology for separation of antitumor active plypeptides from Periplaneta Americana. [Method] Hollow fiber ultrafiltration membrane（3 kDa）was used to separate antitumor active plypeptides from P. Americana. Through single factor test, the impact of rotation speed, concentration ratio, pH and operation time on separation effort was studied, and the separation technology conditions were further optimized through orthogonal test. [Result] The best technology conditions for separation of antitumor active plypeptides from P. Americana were as follows： rotation speed 4 500 r/min, concentration ratio 30%, pH 11, and the rotation speed was the major influencing factor. [Conclusion] The study laid foundation for industrialization of ultrafiltration membrane separation of antitumor active plypeptides from P. Americana.展开更多
Background: The occurrence of bacterial infections sometimes involves synanthropic flies and cockroaches, as mechanical vectors of pathogenic microorganisms. The aim of this study was to identify the different species...Background: The occurrence of bacterial infections sometimes involves synanthropic flies and cockroaches, as mechanical vectors of pathogenic microorganisms. The aim of this study was to identify the different species of flies and cockroaches that cohabit with humans in the city of Cotonou and determine the contribution of Musca domestica and Periplaneta americana in the transmission of bacteria responsible for infections in humans. Methods: Capture sessions during the day for flies and at night for cockroaches were carried out in 4 arrondissements of the city of Cotonou. The insects collected were transported to the Center of Research Entomological of Cotonou for identification, then sent to the Section Hygiene of Water and Food for microbiological analysis, enumeration and testing for human pathogenic bacteria. Results: A total of 351 flies and 26 cockroaches were collected at the sites, including two synanthropic species belonging to two families each. Musca domestica and Periplaneta americana were the most abundant species found after identification in the city of Cotonou. Thermo-tolerant coliforms and Escherichia coli were found on all specimens at varying concentrations. Salmonella spp. bacteria were identified in the fly population from the Dantokpa dump. Conclusion: Flies and cockroaches can transmit microorganisms to humans. The presence of Salmonella spp. among the specimens revealed that infections frequently caused by contaminated food or water are also transmitted to humans by flies. The presence of these germs on flies and cockroaches represents a potential risk of mechanical transmission to humans. It is, therefore, essential to continue investigations in order to assess the species of bacteria propagated, improve control strategies against these troublesome insects and adopt better hygiene conditions for better living.展开更多
The vacuolar (H+)-ATPases (V-ATPases) are ATP-driven proton pumps with multiple functions in many organisms. In this study, we performed structural and functional analysis of vha55 gene that encodes V-ATPase subunit B...The vacuolar (H+)-ATPases (V-ATPases) are ATP-driven proton pumps with multiple functions in many organisms. In this study, we performed structural and functional analysis of vha55 gene that encodes V-ATPase subunit B in the smokybrown cockroach Periplaneta fuliginosa (Blattodea). We observed a high homology score of the deduced amino acid sequences between 10 species in seven orders. RNAi of the vha55 gene in R fuliginosa caused nymphal/nymphal molting defects with incomplete shedding of old cuticles, growth inhibition, as well as bent and wrinkled cuticles of thoraxes and abdominal segments. Since growth inhibition caused by vha55 RNAi did not interfere in the commencement of cockroach molting, molting timing and body growth might be controlled by independent mechanism. Our study suggested V-ATPases might be a good candidate molecule for evolutionary and developmental studies of insect molting.展开更多
The small Rab GTPases are key regulators of membrane vesicle trafficking. Ovaries of Periplaneta americana （Linnaeus） （Blattodea： Blattidae） have small molecular weight GTP/ATP-binding proteins during early and l...The small Rab GTPases are key regulators of membrane vesicle trafficking. Ovaries of Periplaneta americana （Linnaeus） （Blattodea： Blattidae） have small molecular weight GTP/ATP-binding proteins during early and late vitellogenic periods of oogenesis. However, the identification and characterization of the detected proteins have not been yet reported. Herein, we cloned a cDNA encoding Rab5 from the American cockroach, P. americana, ovaries （PamRab5）. It comprises 796 bp, encoding a protein of 213 amino acid residues with a predicted molecular weight of 23.5 kDa. PamRab5 exists as a single-copy gene in the P. americana genome, as revealed by Southern blot analysis. An approximate 2.6 kb ovarian mRNA was transcribed especially at high levels in the previtellogenic ovaries, detected by Northern blot analysis. The muscle and head tissues also showed high levels of PamRab5 transcript. PamRab5 protein was localized, via immunofluorescence labeling, to germline-derived cells of the oocytes, very early during oocyte differentiation. Immunoblotting detected a ~25 kDa signal as a membrane-associated form revealed after application of detergent in the extraction buffer, and 23 kDa as a cytosolic form consistent with the predicted molecular weight from amino acid sequence in different tissues including ovary, muscles and head. The PamRab5 during late vitellogenic periods is required to regulate the endocytotic machinery during oogenesis in this cockroach. This is the first report on Rab5 from a hemimetabolan, and presents an inaugural step in probing the molecular premises of insect oocyte endocytotic trafficking important for oogenesis and embryonic development.展开更多
基金supported by the Natural Science Foundation of Hunan Province(No.2021JJ30516).
文摘Wound healing in diabetic ulcers remains a significant clinical challenge,primarily due to bacterial infection and impaired angiogenesis.Periplaneta americana extract(PAE)has been widely used to treat diabetic wounds,yet its underlying mechanisms are not fully understood.This study aimed to elucidate these mechanisms by analyzing long non-coding RNA(lncRNA)expressions in the wound tissues from diabetic anal fistula patients treated with or without PAE,using high-throughput sequencing.Peripheral blood monocytes from patients were differentiated into M0 macrophages with human macrophage colony-stimulating factor(hMCSF)and subsequently polarized into M1 macrophages with lipopolysaccharide.The results indicated that LINC01133 and SLAMF9 were downregulated in wound tissues of patients treated with PAE.Furthermore,PAE suppressed M1 macrophage polarization and enhanced human umbilical vein endothelial cell(HUVEC)proliferation,migration,and angiogenesis.These effects were diminished when LINC01133 or SLAMF9 were overexpressed.Mechanistically,LINC01133 was shown to upregulate SLAMF9 through interaction with ELAVL1.Overexpression of SLAMF9 reversed the effects of LINC01133 silencing on macrophage polarization and HUVEC functions.In conclusion,PAE facilitates the healing of infected diabetic ulcers by downregulating the LINC01133/SLAMF9 pathway.
基金Supported by the National Natural Science Foundation of China(30560181)the Major Project of Yunnan Province(2008IF012)~~
文摘Researches on pharmacological action and drug development of Periplane- ta americana have demonstrated that it has wide medicinal value, low side effects and rich resources, having good application prospects in medicine field. This article reviewed the researches on pharmacological action and clinical application of Periplaneta americana in order to provide references for the further research and devel- opment of Periplaneta americana medicines.
文摘OBJECTIVE： To investigate the effect of the periplaneta americana extract on the intestinal mucosal barrier and prognostic implications in patients with sepsis. METHODS： Sixty and six patients with sepsis were assigned randomly to treatment group （32 cases） and control group （32 cases）. The extractfrom periplaneta americana plus conventional medication for sepsis was administered to the treatment group, while the control group only received conventional treatment. The gastrointestinal function scores and acute physiology and chronic health evaluation II （APACHE 11 ） scores of all subjects were documented at baseline, at days 1, 3 and 7 after treatment respectively and their blood endotoxin was tested at the same time points as well. The incidence of death was recorded for both groups throughout the trial. RESULTS： At days 3 and 7 after treatment, gastrointestinal function score, APACHE II, and endotoxin level in treatment group wasbetter than that in con- trol group and the difference between them was significant （both P〈0~05）. Although the incidence of death in treatment group was less than that in control group, the difference between the two groups was not significant （P〉 0.05）. CONCLUSION： The extract of periplaneta ameri- cana had protective effect on intestinal mucosal barrier and could improve the condition and prog- nosis in patients with sepsis.
基金supported by Tangshan City Science and Technology Development Project(No.15130219a)
文摘BACKGROUND:Periplaneta americana extract is recognized to have a positive effect on gastrointestinal mucosa.This study aimed to investigate the effects of periplaneta americana extract on immune function,nutrition status and gastrointestinal complications of early enteral nutrition patients with systemic inflammatory response syndrome(SIRS).METHODS:Patients with SIRS were randomly divided into two groups:treatment and control groups.All patients in the two groups received conventional therapy including enteral nutrition,but periplaneta americana extract,an additional Chinese medicine,was given to the patients in the treatment group.At the beginning of treatment(0 day)and 1,3,and 7 days after treatment,the levels of immunoglobulin(Ig A),total lymphocyte count(TLC),total protein(TP)and prealbumin(PA)were respectively tested in patients'venous blood.The incidences of bloating,diarrhea,aspiration pneumonia and high blood sugar at 7 days after treatment were recorded.The mortality of the patients in 28 days was recorded.RESULTS:At 3 and 7 days after treatment,the levels of Ig A and TLC in the treatment group were higher than those in the control group(P<0.05).At 7 days after treatment,the levels of TP and PA in the treatment group were higher than those in the control group(P<0.05).The incidences of bloating and diarrhea in the treatment group were lower than those in the control group,the differences were significant(P<0.05).The mortality of treatment group was lower than that of the control group(P>0.05).CONCLUSION:Periplaneta americana extract could reduce gastrointestinal complications and improve immune function and nutritional status in patients with systemic inflammatory response syndrome.
基金This study was funded in part by the National Natural Science Foundation of China(Nos.31861143050,31772476 and 31911530077).
文摘Objective Acute lung injury(ALI)is an acute clinical syndrome characterized by uncontrolled inflammation response,which causes high mortality and poor prognosis.The present study determined the protective effect and underlying mechanism of Periplaneta americana extract(PAE)against lipopolysaccharide(LPS)-induced ALI.Methods The viability of MH-S cells was measured by MTT.ALI was induced in BALB/c mice by intranasal administration of LPS(5 mg/kg),and the pathological changes,oxidative stress,myeloperoxidase activity,lactate dehydrogenase activity,inflammatory cytokine expression,edema formation,and signal pathway activation in lung tissues and bronchoalveolar lavage fluid(BALF)were examined by H&E staining,MDA,SOD and CAT assays,MPO assay,ELISA,wet/dry analysis,immunofluorescence staining and Western blotting,respectively.Results The results revealed that PAE obviously inhibited the release of proinflammatory TNF-α,IL-6 and IL-1βby suppressing the activation of MAPK/Akt/NF-κB signaling pathways in LPS-treated MH-S cells.Furthermore,PAE suppressed the neutrophil infiltration,permeability increase,pathological changes,cellular damage and death,pro-inflammatory cytokines expression,and oxidative stress upregulation,which was associated with its blockage of the MAPK/Akt/NF-κB pathway in lung tissues of ALI mice.Conclusion PAE may serve as a potential agent for ALI treatment due to its anti-inflammatory and anti-oxidative properties,which correlate to the blockage of the MAPK/NF-κB and AKT signaling pathways.
基金Supported by National Natural Science Foundation of China(30560181)~~
文摘[Objective] The aim of this study was to investigate the decolorization ef- fects of activated carbon and macroporous resin on Periplaneta americana L. skimmed cream and compare the advantages and disadvantages of the two decolorization technologies. [Method] Periplaneta americana L. skimmed cream was decolored with activated carbon and macroporous resin, and freeze-dried to collect solid decolorization products. By investigating the yield, decolorization rate, protein retention rate and decolorization operation process, the advantages and disadvantages of the two decolorization technologies were compared. [Result] Both activated carbon and macroporous resin can be used for decolorization. To be specific, macroporous resin-decolorization is superior in the yield and protein retention rate, while activated carbon-decolorization is superior in decolorization rate and decolorization operation process. [Conclusion] Macroporous resin-decolorization can be used if protein is the main ingredient required in the experiment, while activated carbon-decolorization can be used if protein is not the main ingredient required.
基金Supported by National Natural Science Foundation of China ( 30560181)Key Industry Innovation Project of Yunnan Province ( 2008IF012)
文摘[ Objective] This study aimed to explore a Tricine-SDS-PAGE method for separation and purification of Periplaneta Americana small peptides. [ Method] By comparing the separation effects of three types of gels with different ratios on P. Americana small peptides, an appropriate Tricine-SDS-PAGE method for separation of small peptides was established. [ Result] Standard low molecular weight Marker was preliminarily separated to a certain extent with polyacrylamide gel electraphoresis, while the sample mixture was not separated due to the different levels of aggregation in separating gel, spacer gel and stacking gel. [Condusion] More appropriate polyacrylamide gel electrephoresis separation method or other better separation methods were to be further explored for separating peptide mixture with smaller molecular weight.
基金Supported by Fund Project of Key Industrial Innovation Engineering of Yunnan Province(2008IF012)
文摘[Objective] The paper was to establish ultrafiltration membrane technology for separation of antitumor active plypeptides from Periplaneta Americana. [Method] Hollow fiber ultrafiltration membrane（3 kDa）was used to separate antitumor active plypeptides from P. Americana. Through single factor test, the impact of rotation speed, concentration ratio, pH and operation time on separation effort was studied, and the separation technology conditions were further optimized through orthogonal test. [Result] The best technology conditions for separation of antitumor active plypeptides from P. Americana were as follows： rotation speed 4 500 r/min, concentration ratio 30%, pH 11, and the rotation speed was the major influencing factor. [Conclusion] The study laid foundation for industrialization of ultrafiltration membrane separation of antitumor active plypeptides from P. Americana.
文摘Background: The occurrence of bacterial infections sometimes involves synanthropic flies and cockroaches, as mechanical vectors of pathogenic microorganisms. The aim of this study was to identify the different species of flies and cockroaches that cohabit with humans in the city of Cotonou and determine the contribution of Musca domestica and Periplaneta americana in the transmission of bacteria responsible for infections in humans. Methods: Capture sessions during the day for flies and at night for cockroaches were carried out in 4 arrondissements of the city of Cotonou. The insects collected were transported to the Center of Research Entomological of Cotonou for identification, then sent to the Section Hygiene of Water and Food for microbiological analysis, enumeration and testing for human pathogenic bacteria. Results: A total of 351 flies and 26 cockroaches were collected at the sites, including two synanthropic species belonging to two families each. Musca domestica and Periplaneta americana were the most abundant species found after identification in the city of Cotonou. Thermo-tolerant coliforms and Escherichia coli were found on all specimens at varying concentrations. Salmonella spp. bacteria were identified in the fly population from the Dantokpa dump. Conclusion: Flies and cockroaches can transmit microorganisms to humans. The presence of Salmonella spp. among the specimens revealed that infections frequently caused by contaminated food or water are also transmitted to humans by flies. The presence of these germs on flies and cockroaches represents a potential risk of mechanical transmission to humans. It is, therefore, essential to continue investigations in order to assess the species of bacteria propagated, improve control strategies against these troublesome insects and adopt better hygiene conditions for better living.
文摘The vacuolar (H+)-ATPases (V-ATPases) are ATP-driven proton pumps with multiple functions in many organisms. In this study, we performed structural and functional analysis of vha55 gene that encodes V-ATPase subunit B in the smokybrown cockroach Periplaneta fuliginosa (Blattodea). We observed a high homology score of the deduced amino acid sequences between 10 species in seven orders. RNAi of the vha55 gene in R fuliginosa caused nymphal/nymphal molting defects with incomplete shedding of old cuticles, growth inhibition, as well as bent and wrinkled cuticles of thoraxes and abdominal segments. Since growth inhibition caused by vha55 RNAi did not interfere in the commencement of cockroach molting, molting timing and body growth might be controlled by independent mechanism. Our study suggested V-ATPases might be a good candidate molecule for evolutionary and developmental studies of insect molting.
文摘The small Rab GTPases are key regulators of membrane vesicle trafficking. Ovaries of Periplaneta americana （Linnaeus） （Blattodea： Blattidae） have small molecular weight GTP/ATP-binding proteins during early and late vitellogenic periods of oogenesis. However, the identification and characterization of the detected proteins have not been yet reported. Herein, we cloned a cDNA encoding Rab5 from the American cockroach, P. americana, ovaries （PamRab5）. It comprises 796 bp, encoding a protein of 213 amino acid residues with a predicted molecular weight of 23.5 kDa. PamRab5 exists as a single-copy gene in the P. americana genome, as revealed by Southern blot analysis. An approximate 2.6 kb ovarian mRNA was transcribed especially at high levels in the previtellogenic ovaries, detected by Northern blot analysis. The muscle and head tissues also showed high levels of PamRab5 transcript. PamRab5 protein was localized, via immunofluorescence labeling, to germline-derived cells of the oocytes, very early during oocyte differentiation. Immunoblotting detected a ~25 kDa signal as a membrane-associated form revealed after application of detergent in the extraction buffer, and 23 kDa as a cytosolic form consistent with the predicted molecular weight from amino acid sequence in different tissues including ovary, muscles and head. The PamRab5 during late vitellogenic periods is required to regulate the endocytotic machinery during oogenesis in this cockroach. This is the first report on Rab5 from a hemimetabolan, and presents an inaugural step in probing the molecular premises of insect oocyte endocytotic trafficking important for oogenesis and embryonic development.
