Susilorini has completed her Master of Medical Science at the age of 30 years from Magister of Biomedic of Diponegoro University and Pathology Anatomy Residency from Medical faculty of Diponegoro University. She is a pathologist and staff of Departement Pathology Anatomy of Medical faculty of Sultan Agung Islamic University and Sultan Agung Islamic hospital. Her research foccused in the prophetic medicine.
Breast milk is a complex liquid with immune-competent cells and soluble proteins that provide imm... more Breast milk is a complex liquid with immune-competent cells and soluble proteins that provide immunity to the infant and affect
the maturation of the infant’s immune system. Exosomes are nanovesicles (30–100 nm) with an endosome-derived limiting
membrane secreted by a diverse range of cell types. Because exosomes carry immunorelevant structures, they are suggested to
participate in directing the immune response. We hypothesized that human breast milk contain exosomes, which may be important
for the development of the infant’s immune system. We isolated vesicles from the human colostrum and mature breast milk by
ultracentrifugations and/or immuno-isolation on paramagnetic beads. We found that the vesicles displayed a typical exosome-like
size and morphology as analyzed by electron microscopy. Furthermore, they floated at a density between 1.10 and 1.18 g/ml in a
sucrose gradient, corresponding to the known density of exosomes. In addition, MHC classes I and II, CD63, CD81, and CD86 were
detected on the vesicles by flow cytometry. Western blot and mass spectrometry further confirmed the presence of several
exosome-associated molecules. Functional analysis revealed that the vesicle preparation inhibited anti-CD3-induced IL-2 and
IFN- production from allogeneic and autologous PBMC. In addition, an increased number of Foxp3CD4CD25 T regulatory
cells were observed in PBMC incubated with milk vesicle preparations. We conclude that human breast milk contains exosomes
with the capacity to influence immune responses. The Journal of Immunology, 2007, 179: 1969–1978.
The release of exosomes and other microvesicles
by diverse prokaryotic and eukaryotic cells and
o... more The release of exosomes and other microvesicles by diverse prokaryotic and eukaryotic cells and organisms was first appreciated early in the 20th century. The functional properties of these organelles, however, have only recently been the focus of rigorous investigation. In this review, we discuss the release of microvesicles of varying complexity by diverse microbial pathogens. This includes vesicle secretion by Gram-negative bacteria, eukaryotic parasites of the kinetoplast lineage and opportunistic fungal pathogens of both the ascomycetes and basidiomycetes lineages. We also discuss vesicle release from mammalian cells brought about as a result of infection with bacteria, viruses and prions. In addition, we review the evidence showing that in their specific microenvironments, release of these organelles from diverse pathogens contributes to pathogenesis. Germane to this and based upon recent findings with Leishmania, we propose a model whereby exosome release by an intracellular pathogen serves as a general mechanism for effector molecule delivery from eukaryotic pathogen to host cell cytosol. These new findings linking exosomes and other microvesicles to infection biology have important implications for understanding the immune response to infection and for the design of research strategies aimed at the development of novel therapeutics and vaccines
Abstract
Background: Exosomes are 30-100 nm membrane vesicles of endocytic origin produced by num... more Abstract Background: Exosomes are 30-100 nm membrane vesicles of endocytic origin produced by numerous cells. They can mediate diverse biological functions, including antigen presentation. Exosomes have recently been shown to contain functional RNA, which can be delivered to other cells. Exosomes may thus mediate biological functions either by surface-to-surface interactions with cells, or by the delivery of functional RNA to cells. Our aim was therefore to determine the presence of RNA in exosomes from human saliva, plasma and breast milk and whether these exosomes can be taken up by macrophages. Method: Exosomes were purified from human saliva, plasma and breast milk using ultracentrifugation and filtration steps. Exosomes were detected by electron microscopy and examined by flow cytometry. Flow cytometry was performed by capturing the exosomes on anti-MHC class II coated beads, and further stain with anti-CD9, anti- CD63 or anti-CD81. Breast milk exosomes were further analysed for the presence of Hsc70, CD81 and calnexin by Western blot. Total RNA was detected with a Bioanalyzer and mRNA was identified by the synthesis of cDNA using an oligo (dT) primer and analysed with a Bioanalyzer. The uptake of PKH67-labelled saliva and breast milk exosomes by macrophages was examined by measuring fluorescence using flow cytometry and fluorescence microscopy. Results: RNA was detected in exosomes from all three body fluids. A portion of the detected RNA in plasma exosomes was characterised as mRNA. Our result extends the characterisation of exosomes in healthy humans and confirms the presence of RNA in human saliva and plasma exosomes and reports for the first time the presence of RNA in breast milk exosomes. Our results also show that the saliva and breast milk exosomes can be taken up by human macrophages. Conclusions: Exosomes in saliva, plasma and breast milk all contain RNA, confirming previous findings that exosomes from several sources contain RNA. Furthermore, exosomes are readily taken up by macrophages, supporting the notion that exosomal RNA can be shuttled between cells
Background
Cupping is a therapeutic technique employed by many acupuncturists to treat Full,
Mixe... more Background Cupping is a therapeutic technique employed by many acupuncturists to treat Full, Mixed or Deficiency conditions. Despite its documented effectiveness, however, little is known from a Western scientific standpoint about the mechanisms by which cupping therapy yields its effects. In 2005, Ilkay Chirali published the results of a small trial conducted on 6 patients with musculoskeletal complaints. In this study patients were treated every two days over a two weekly period and blood samples were taken and analysed before, during and after the treatment period. Analysis of the samples revealed a significant reduction in the Erythrocyte Sedimentation Rate (ESR), a broad marker of inflammation. The aim of the current study was to identify inflammatory marker(s) whose plasma concentrations are altered during the cupping therapy in the treatment of musculoskeletal disorders and which may account for changes in the ESR. This will provide biochemical evidence for the effectiveness of cupping therapy. Methodology 14 subjects presenting with a range of chronic musculoskeletal complaints (diagnosed as Empty, Mixed and Full conditions) were recruited into the study. Full informed consent was obtained from each subject. A Traditional Diagnosis was then performed on each subject and the cupping treatment strategy determined accordingly. Each subject then received six cupping treatments at weekly intervals. Prior to the first treatment and thereafter at weekly intervals, subjects completed a MYMOP questionnaire in order to assess clinical outcome. Venous blood samples were also obtained before treatment, after 3 treatments, after 6 treatments and six weeks post treatment. The following analyses were performed on blood samples: full blood count, haemoglobin concentration, erythrocyte sedimentation rate (ESR), serum concentration of fibrinogen, C Reactive Protein (CRP), IgG and ferritin. Serum samples were also analysed for the concentration of inflammatory cytokines (IL-1β, IL-6, IL-10 and TNF-α). Data obtained was analysed by one way ANOVA with matched values and Dunnett’s Multiple Comparisons Test using Instat2 software. Results 9 females (range 23 – 52 years) and 5 males (range 37 – 62 years) were recruited into the study. One female participant subsequently withdrew from the study at week 3. Analysis of MYMOP questionnaires indicated that 95% patients reported improvements in their symptoms as a result of treatment and, overall, this improvement was found to significant. Complete data sets from blood and serum analysis (before, during and after treatment) were obtained for 11 subjects. The six week post treatment follow up was 54%. Analysis of data revealed statistically significant increases in the platelet count (p = 0.0373) and lymphocyte count (p = 0.0001) and decreases in the serum concentration of fibrinogen (p = 0.0008) and ferritin (p = 0.0024). No changes were measured in the serum concentration of proinflammatory cytokines, with the exception of one patient where slight increases above normal levels were seen in TNF-α, IL-6 and IL-10 concentrations. Interestingly there was no significant change in the ESR observed in these subjects, however, only 3 subjects presented with an elevated ESR before treatment. Discussion & Conclusion Ferritin and fibrinogen are inflammatory markers and their reduction during cupping therapy is concomitant with a reduction in the inflammatory status of the patient. Platelets and lymphocytes may increase as a result of local vascular damage from the cupping therapy itself or from a reduction in the adherence of these cells to areas of activated endothelium, associated with inflammation. With regard to the ESR, no overall significant change was observed in this study. However, only 3 patients presented with an elevated ESR prior to treatment. Furthermore, the ESR did decrease during the course of treatment in these subjects. In conclusion, although the precise mechanism by which cupping therapy exerts it therapeutic effects cannot be determined from the present study, the results indicate that cupping therapy is associated with significant changes in the levels of inflammatory cells and soluble markers suggesting that this treatment can influence the inflammatory status of the patient leading to improved clinical outcome
Abstract
A number of chemicals have been shown to demonstrate neurotoxic effects either in human ... more Abstract A number of chemicals have been shown to demonstrate neurotoxic effects either in human or laboratory animal studies. This article aims at evaluating the impact of exposure to several chemicals including: organophosphate, organochlorine pesticides, polychlorinated biphenyls (PCBs), mercury and lead on the neurodevelopment of children by reviewing the most recent published literature, and answer the question whether any progress has been made in the epidemiology of the neurodevelopment of children induced by exposure to those chemicals. The result of the presented studies show that exposure to the above-mentioned chemicals may impair the neurodevelopment of children. Neonates exposed to organophosphate pesticides demonstrated a higher proportion of abnormal reflexes, and young children had more attention problems. Exposure to organochlorine pesticides in children was associated with alertness, quality of alert responsiveness, cost of attention and other potential attention associated measures. The majority of studies indicate the negative impact of lead exposure at the level <10 μg/dl or even <5 μg/dl on the neurodevelopment of children. The results of studies on exposure to PCBs, mercury, and their effect on neurodevelopment are inconsistent. Some suggest that prenatal exposure to PCBs and mercury is related to performance impairments, attention and concentration problems, while other do not present any statistically significant association. The studies were mostly well designed, using prospective cohorts with the exposure assessment based on the biomarker of exposure. Concerning the covariates and confounders affecting the endpoints in most of the presented studies, confounders were included in data analysis. In order to recognize the early cognitive, motor and language outcomes of chemical exposures, well standardized tools were used for evaluating the neurodevelopmental effects and offer an early and fairly comprehensive measure of child development. Because the neurotoxicants may cross the placenta and the fetal brain, exposure consideration regarding the reduction of exposure to those chemicals should be implemented. Key words pesticides, PCB, mercury, lead, neurodevelopment, exposure assessment
Breast milk is a complex liquid with immune-competent cells and soluble proteins that provide imm... more Breast milk is a complex liquid with immune-competent cells and soluble proteins that provide immunity to the infant and affect
the maturation of the infant’s immune system. Exosomes are nanovesicles (30–100 nm) with an endosome-derived limiting
membrane secreted by a diverse range of cell types. Because exosomes carry immunorelevant structures, they are suggested to
participate in directing the immune response. We hypothesized that human breast milk contain exosomes, which may be important
for the development of the infant’s immune system. We isolated vesicles from the human colostrum and mature breast milk by
ultracentrifugations and/or immuno-isolation on paramagnetic beads. We found that the vesicles displayed a typical exosome-like
size and morphology as analyzed by electron microscopy. Furthermore, they floated at a density between 1.10 and 1.18 g/ml in a
sucrose gradient, corresponding to the known density of exosomes. In addition, MHC classes I and II, CD63, CD81, and CD86 were
detected on the vesicles by flow cytometry. Western blot and mass spectrometry further confirmed the presence of several
exosome-associated molecules. Functional analysis revealed that the vesicle preparation inhibited anti-CD3-induced IL-2 and
IFN- production from allogeneic and autologous PBMC. In addition, an increased number of Foxp3CD4CD25 T regulatory
cells were observed in PBMC incubated with milk vesicle preparations. We conclude that human breast milk contains exosomes
with the capacity to influence immune responses. The Journal of Immunology, 2007, 179: 1969–1978.
The release of exosomes and other microvesicles
by diverse prokaryotic and eukaryotic cells and
o... more The release of exosomes and other microvesicles by diverse prokaryotic and eukaryotic cells and organisms was first appreciated early in the 20th century. The functional properties of these organelles, however, have only recently been the focus of rigorous investigation. In this review, we discuss the release of microvesicles of varying complexity by diverse microbial pathogens. This includes vesicle secretion by Gram-negative bacteria, eukaryotic parasites of the kinetoplast lineage and opportunistic fungal pathogens of both the ascomycetes and basidiomycetes lineages. We also discuss vesicle release from mammalian cells brought about as a result of infection with bacteria, viruses and prions. In addition, we review the evidence showing that in their specific microenvironments, release of these organelles from diverse pathogens contributes to pathogenesis. Germane to this and based upon recent findings with Leishmania, we propose a model whereby exosome release by an intracellular pathogen serves as a general mechanism for effector molecule delivery from eukaryotic pathogen to host cell cytosol. These new findings linking exosomes and other microvesicles to infection biology have important implications for understanding the immune response to infection and for the design of research strategies aimed at the development of novel therapeutics and vaccines
Abstract
Background: Exosomes are 30-100 nm membrane vesicles of endocytic origin produced by num... more Abstract Background: Exosomes are 30-100 nm membrane vesicles of endocytic origin produced by numerous cells. They can mediate diverse biological functions, including antigen presentation. Exosomes have recently been shown to contain functional RNA, which can be delivered to other cells. Exosomes may thus mediate biological functions either by surface-to-surface interactions with cells, or by the delivery of functional RNA to cells. Our aim was therefore to determine the presence of RNA in exosomes from human saliva, plasma and breast milk and whether these exosomes can be taken up by macrophages. Method: Exosomes were purified from human saliva, plasma and breast milk using ultracentrifugation and filtration steps. Exosomes were detected by electron microscopy and examined by flow cytometry. Flow cytometry was performed by capturing the exosomes on anti-MHC class II coated beads, and further stain with anti-CD9, anti- CD63 or anti-CD81. Breast milk exosomes were further analysed for the presence of Hsc70, CD81 and calnexin by Western blot. Total RNA was detected with a Bioanalyzer and mRNA was identified by the synthesis of cDNA using an oligo (dT) primer and analysed with a Bioanalyzer. The uptake of PKH67-labelled saliva and breast milk exosomes by macrophages was examined by measuring fluorescence using flow cytometry and fluorescence microscopy. Results: RNA was detected in exosomes from all three body fluids. A portion of the detected RNA in plasma exosomes was characterised as mRNA. Our result extends the characterisation of exosomes in healthy humans and confirms the presence of RNA in human saliva and plasma exosomes and reports for the first time the presence of RNA in breast milk exosomes. Our results also show that the saliva and breast milk exosomes can be taken up by human macrophages. Conclusions: Exosomes in saliva, plasma and breast milk all contain RNA, confirming previous findings that exosomes from several sources contain RNA. Furthermore, exosomes are readily taken up by macrophages, supporting the notion that exosomal RNA can be shuttled between cells
Background
Cupping is a therapeutic technique employed by many acupuncturists to treat Full,
Mixe... more Background Cupping is a therapeutic technique employed by many acupuncturists to treat Full, Mixed or Deficiency conditions. Despite its documented effectiveness, however, little is known from a Western scientific standpoint about the mechanisms by which cupping therapy yields its effects. In 2005, Ilkay Chirali published the results of a small trial conducted on 6 patients with musculoskeletal complaints. In this study patients were treated every two days over a two weekly period and blood samples were taken and analysed before, during and after the treatment period. Analysis of the samples revealed a significant reduction in the Erythrocyte Sedimentation Rate (ESR), a broad marker of inflammation. The aim of the current study was to identify inflammatory marker(s) whose plasma concentrations are altered during the cupping therapy in the treatment of musculoskeletal disorders and which may account for changes in the ESR. This will provide biochemical evidence for the effectiveness of cupping therapy. Methodology 14 subjects presenting with a range of chronic musculoskeletal complaints (diagnosed as Empty, Mixed and Full conditions) were recruited into the study. Full informed consent was obtained from each subject. A Traditional Diagnosis was then performed on each subject and the cupping treatment strategy determined accordingly. Each subject then received six cupping treatments at weekly intervals. Prior to the first treatment and thereafter at weekly intervals, subjects completed a MYMOP questionnaire in order to assess clinical outcome. Venous blood samples were also obtained before treatment, after 3 treatments, after 6 treatments and six weeks post treatment. The following analyses were performed on blood samples: full blood count, haemoglobin concentration, erythrocyte sedimentation rate (ESR), serum concentration of fibrinogen, C Reactive Protein (CRP), IgG and ferritin. Serum samples were also analysed for the concentration of inflammatory cytokines (IL-1β, IL-6, IL-10 and TNF-α). Data obtained was analysed by one way ANOVA with matched values and Dunnett’s Multiple Comparisons Test using Instat2 software. Results 9 females (range 23 – 52 years) and 5 males (range 37 – 62 years) were recruited into the study. One female participant subsequently withdrew from the study at week 3. Analysis of MYMOP questionnaires indicated that 95% patients reported improvements in their symptoms as a result of treatment and, overall, this improvement was found to significant. Complete data sets from blood and serum analysis (before, during and after treatment) were obtained for 11 subjects. The six week post treatment follow up was 54%. Analysis of data revealed statistically significant increases in the platelet count (p = 0.0373) and lymphocyte count (p = 0.0001) and decreases in the serum concentration of fibrinogen (p = 0.0008) and ferritin (p = 0.0024). No changes were measured in the serum concentration of proinflammatory cytokines, with the exception of one patient where slight increases above normal levels were seen in TNF-α, IL-6 and IL-10 concentrations. Interestingly there was no significant change in the ESR observed in these subjects, however, only 3 subjects presented with an elevated ESR before treatment. Discussion & Conclusion Ferritin and fibrinogen are inflammatory markers and their reduction during cupping therapy is concomitant with a reduction in the inflammatory status of the patient. Platelets and lymphocytes may increase as a result of local vascular damage from the cupping therapy itself or from a reduction in the adherence of these cells to areas of activated endothelium, associated with inflammation. With regard to the ESR, no overall significant change was observed in this study. However, only 3 patients presented with an elevated ESR prior to treatment. Furthermore, the ESR did decrease during the course of treatment in these subjects. In conclusion, although the precise mechanism by which cupping therapy exerts it therapeutic effects cannot be determined from the present study, the results indicate that cupping therapy is associated with significant changes in the levels of inflammatory cells and soluble markers suggesting that this treatment can influence the inflammatory status of the patient leading to improved clinical outcome
Abstract
A number of chemicals have been shown to demonstrate neurotoxic effects either in human ... more Abstract A number of chemicals have been shown to demonstrate neurotoxic effects either in human or laboratory animal studies. This article aims at evaluating the impact of exposure to several chemicals including: organophosphate, organochlorine pesticides, polychlorinated biphenyls (PCBs), mercury and lead on the neurodevelopment of children by reviewing the most recent published literature, and answer the question whether any progress has been made in the epidemiology of the neurodevelopment of children induced by exposure to those chemicals. The result of the presented studies show that exposure to the above-mentioned chemicals may impair the neurodevelopment of children. Neonates exposed to organophosphate pesticides demonstrated a higher proportion of abnormal reflexes, and young children had more attention problems. Exposure to organochlorine pesticides in children was associated with alertness, quality of alert responsiveness, cost of attention and other potential attention associated measures. The majority of studies indicate the negative impact of lead exposure at the level <10 μg/dl or even <5 μg/dl on the neurodevelopment of children. The results of studies on exposure to PCBs, mercury, and their effect on neurodevelopment are inconsistent. Some suggest that prenatal exposure to PCBs and mercury is related to performance impairments, attention and concentration problems, while other do not present any statistically significant association. The studies were mostly well designed, using prospective cohorts with the exposure assessment based on the biomarker of exposure. Concerning the covariates and confounders affecting the endpoints in most of the presented studies, confounders were included in data analysis. In order to recognize the early cognitive, motor and language outcomes of chemical exposures, well standardized tools were used for evaluating the neurodevelopmental effects and offer an early and fairly comprehensive measure of child development. Because the neurotoxicants may cross the placenta and the fetal brain, exposure consideration regarding the reduction of exposure to those chemicals should be implemented. Key words pesticides, PCB, mercury, lead, neurodevelopment, exposure assessment
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Papers by susilorini sukamto
the maturation of the infant’s immune system. Exosomes are nanovesicles (30–100 nm) with an endosome-derived limiting
membrane secreted by a diverse range of cell types. Because exosomes carry immunorelevant structures, they are suggested to
participate in directing the immune response. We hypothesized that human breast milk contain exosomes, which may be important
for the development of the infant’s immune system. We isolated vesicles from the human colostrum and mature breast milk by
ultracentrifugations and/or immuno-isolation on paramagnetic beads. We found that the vesicles displayed a typical exosome-like
size and morphology as analyzed by electron microscopy. Furthermore, they floated at a density between 1.10 and 1.18 g/ml in a
sucrose gradient, corresponding to the known density of exosomes. In addition, MHC classes I and II, CD63, CD81, and CD86 were
detected on the vesicles by flow cytometry. Western blot and mass spectrometry further confirmed the presence of several
exosome-associated molecules. Functional analysis revealed that the vesicle preparation inhibited anti-CD3-induced IL-2 and
IFN- production from allogeneic and autologous PBMC. In addition, an increased number of Foxp3CD4CD25 T regulatory
cells were observed in PBMC incubated with milk vesicle preparations. We conclude that human breast milk contains exosomes
with the capacity to influence immune responses. The Journal of Immunology, 2007, 179: 1969–1978.
by diverse prokaryotic and eukaryotic cells and
organisms was first appreciated early in the
20th century. The functional properties of these
organelles, however, have only recently been the
focus of rigorous investigation. In this review, we
discuss the release of microvesicles of varying
complexity by diverse microbial pathogens. This
includes vesicle secretion by Gram-negative bacteria,
eukaryotic parasites of the kinetoplast
lineage and opportunistic fungal pathogens of
both the ascomycetes and basidiomycetes lineages.
We also discuss vesicle release from mammalian
cells brought about as a result of infection
with bacteria, viruses and prions. In addition, we
review the evidence showing that in their specific
microenvironments, release of these organelles
from diverse pathogens contributes to pathogenesis.
Germane to this and based upon recent
findings with Leishmania, we propose a model
whereby exosome release by an intracellular
pathogen serves as a general mechanism for effector
molecule delivery from eukaryotic pathogen
to host cell cytosol. These new findings linking
exosomes and other microvesicles to infection
biology have important implications for understanding
the immune response to infection and for
the design of research strategies aimed at the
development of novel therapeutics and vaccines
Background: Exosomes are 30-100 nm membrane vesicles of endocytic origin produced by numerous cells. They
can mediate diverse biological functions, including antigen presentation. Exosomes have recently been shown to
contain functional RNA, which can be delivered to other cells. Exosomes may thus mediate biological functions
either by surface-to-surface interactions with cells, or by the delivery of functional RNA to cells. Our aim was
therefore to determine the presence of RNA in exosomes from human saliva, plasma and breast milk and whether
these exosomes can be taken up by macrophages.
Method: Exosomes were purified from human saliva, plasma and breast milk using ultracentrifugation and filtration
steps. Exosomes were detected by electron microscopy and examined by flow cytometry. Flow cytometry was
performed by capturing the exosomes on anti-MHC class II coated beads, and further stain with anti-CD9, anti-
CD63 or anti-CD81. Breast milk exosomes were further analysed for the presence of Hsc70, CD81 and calnexin by
Western blot. Total RNA was detected with a Bioanalyzer and mRNA was identified by the synthesis of cDNA using
an oligo (dT) primer and analysed with a Bioanalyzer. The uptake of PKH67-labelled saliva and breast milk
exosomes by macrophages was examined by measuring fluorescence using flow cytometry and fluorescence
microscopy.
Results: RNA was detected in exosomes from all three body fluids. A portion of the detected RNA in plasma
exosomes was characterised as mRNA. Our result extends the characterisation of exosomes in healthy humans and
confirms the presence of RNA in human saliva and plasma exosomes and reports for the first time the presence of
RNA in breast milk exosomes. Our results also show that the saliva and breast milk exosomes can be taken up by
human macrophages.
Conclusions: Exosomes in saliva, plasma and breast milk all contain RNA, confirming previous findings that
exosomes from several sources contain RNA. Furthermore, exosomes are readily taken up by macrophages,
supporting the notion that exosomal RNA can be shuttled between cells
Cupping is a therapeutic technique employed by many acupuncturists to treat Full,
Mixed or Deficiency conditions. Despite its documented effectiveness, however, little
is known from a Western scientific standpoint about the mechanisms by which
cupping therapy yields its effects. In 2005, Ilkay Chirali published the results of a
small trial conducted on 6 patients with musculoskeletal complaints. In this study
patients were treated every two days over a two weekly period and blood samples
were taken and analysed before, during and after the treatment period. Analysis of the
samples revealed a significant reduction in the Erythrocyte Sedimentation Rate
(ESR), a broad marker of inflammation. The aim of the current study was to identify
inflammatory marker(s) whose plasma concentrations are altered during the cupping
therapy in the treatment of musculoskeletal disorders and which may account for
changes in the ESR. This will provide biochemical evidence for the effectiveness of
cupping therapy.
Methodology
14 subjects presenting with a range of chronic musculoskeletal complaints (diagnosed
as Empty, Mixed and Full conditions) were recruited into the study. Full informed
consent was obtained from each subject. A Traditional Diagnosis was then performed
on each subject and the cupping treatment strategy determined accordingly. Each
subject then received six cupping treatments at weekly intervals. Prior to the first
treatment and thereafter at weekly intervals, subjects completed a MYMOP
questionnaire in order to assess clinical outcome. Venous blood samples were also
obtained before treatment, after 3 treatments, after 6 treatments and six weeks post
treatment. The following analyses were performed on blood samples: full blood
count, haemoglobin concentration, erythrocyte sedimentation rate (ESR), serum
concentration of fibrinogen, C Reactive Protein (CRP), IgG and ferritin. Serum
samples were also analysed for the concentration of inflammatory cytokines (IL-1β,
IL-6, IL-10 and TNF-α). Data obtained was analysed by one way ANOVA with
matched values and Dunnett’s Multiple Comparisons Test using Instat2 software.
Results
9 females (range 23 – 52 years) and 5 males (range 37 – 62 years) were recruited into
the study. One female participant subsequently withdrew from the study at week 3.
Analysis of MYMOP questionnaires indicated that 95% patients reported
improvements in their symptoms as a result of treatment and, overall, this
improvement was found to significant.
Complete data sets from blood and serum analysis (before, during and after treatment)
were obtained for 11 subjects. The six week post treatment follow up was 54%.
Analysis of data revealed statistically significant increases in the platelet count (p =
0.0373) and lymphocyte count (p = 0.0001) and decreases in the serum concentration
of fibrinogen (p = 0.0008) and ferritin (p = 0.0024). No changes were measured in the
serum concentration of proinflammatory cytokines, with the exception of one patient
where slight increases above normal levels were seen in TNF-α, IL-6 and IL-10
concentrations. Interestingly there was no significant change in the ESR observed in
these subjects, however, only 3 subjects presented with an elevated ESR before
treatment.
Discussion & Conclusion
Ferritin and fibrinogen are inflammatory markers and their reduction during cupping
therapy is concomitant with a reduction in the inflammatory status of the patient.
Platelets and lymphocytes may increase as a result of local vascular damage from the
cupping therapy itself or from a reduction in the adherence of these cells to areas of
activated endothelium, associated with inflammation. With regard to the ESR, no
overall significant change was observed in this study. However, only 3 patients
presented with an elevated ESR prior to treatment. Furthermore, the ESR did decrease
during the course of treatment in these subjects. In conclusion, although the precise
mechanism by which cupping therapy exerts it therapeutic effects cannot be
determined from the present study, the results indicate that cupping therapy is
associated with significant changes in the levels of inflammatory cells and soluble
markers suggesting that this treatment can influence the inflammatory status of the
patient leading to improved clinical outcome
A number of chemicals have been shown to demonstrate neurotoxic effects either in human or laboratory animal studies.
This article aims at evaluating the impact of exposure to several chemicals including: organophosphate, organochlorine
pesticides, polychlorinated biphenyls (PCBs), mercury and lead on the neurodevelopment of children by reviewing the
most recent published literature, and answer the question whether any progress has been made in the epidemiology
of the neurodevelopment of children induced by exposure to those chemicals. The result of the presented studies show
that exposure to the above-mentioned chemicals may impair the neurodevelopment of children. Neonates exposed to
organophosphate pesticides demonstrated a higher proportion of abnormal reflexes, and young children had more attention
problems. Exposure to organochlorine pesticides in children was associated with alertness, quality of alert responsiveness,
cost of attention and other potential attention associated measures. The majority of studies indicate the negative impact
of lead exposure at the level <10 μg/dl or even <5 μg/dl on the neurodevelopment of children. The results of studies on
exposure to PCBs, mercury, and their effect on neurodevelopment are inconsistent. Some suggest that prenatal exposure
to PCBs and mercury is related to performance impairments, attention and concentration problems, while other do not
present any statistically significant association. The studies were mostly well designed, using prospective cohorts with
the exposure assessment based on the biomarker of exposure. Concerning the covariates and confounders affecting the
endpoints in most of the presented studies, confounders were included in data analysis. In order to recognize the early
cognitive, motor and language outcomes of chemical exposures, well standardized tools were used for evaluating the
neurodevelopmental effects and offer an early and fairly comprehensive measure of child development. Because the
neurotoxicants may cross the placenta and the fetal brain, exposure consideration regarding the reduction of exposure to
those chemicals should be implemented.
Key words
pesticides, PCB, mercury, lead, neurodevelopment, exposure assessment
the maturation of the infant’s immune system. Exosomes are nanovesicles (30–100 nm) with an endosome-derived limiting
membrane secreted by a diverse range of cell types. Because exosomes carry immunorelevant structures, they are suggested to
participate in directing the immune response. We hypothesized that human breast milk contain exosomes, which may be important
for the development of the infant’s immune system. We isolated vesicles from the human colostrum and mature breast milk by
ultracentrifugations and/or immuno-isolation on paramagnetic beads. We found that the vesicles displayed a typical exosome-like
size and morphology as analyzed by electron microscopy. Furthermore, they floated at a density between 1.10 and 1.18 g/ml in a
sucrose gradient, corresponding to the known density of exosomes. In addition, MHC classes I and II, CD63, CD81, and CD86 were
detected on the vesicles by flow cytometry. Western blot and mass spectrometry further confirmed the presence of several
exosome-associated molecules. Functional analysis revealed that the vesicle preparation inhibited anti-CD3-induced IL-2 and
IFN- production from allogeneic and autologous PBMC. In addition, an increased number of Foxp3CD4CD25 T regulatory
cells were observed in PBMC incubated with milk vesicle preparations. We conclude that human breast milk contains exosomes
with the capacity to influence immune responses. The Journal of Immunology, 2007, 179: 1969–1978.
by diverse prokaryotic and eukaryotic cells and
organisms was first appreciated early in the
20th century. The functional properties of these
organelles, however, have only recently been the
focus of rigorous investigation. In this review, we
discuss the release of microvesicles of varying
complexity by diverse microbial pathogens. This
includes vesicle secretion by Gram-negative bacteria,
eukaryotic parasites of the kinetoplast
lineage and opportunistic fungal pathogens of
both the ascomycetes and basidiomycetes lineages.
We also discuss vesicle release from mammalian
cells brought about as a result of infection
with bacteria, viruses and prions. In addition, we
review the evidence showing that in their specific
microenvironments, release of these organelles
from diverse pathogens contributes to pathogenesis.
Germane to this and based upon recent
findings with Leishmania, we propose a model
whereby exosome release by an intracellular
pathogen serves as a general mechanism for effector
molecule delivery from eukaryotic pathogen
to host cell cytosol. These new findings linking
exosomes and other microvesicles to infection
biology have important implications for understanding
the immune response to infection and for
the design of research strategies aimed at the
development of novel therapeutics and vaccines
Background: Exosomes are 30-100 nm membrane vesicles of endocytic origin produced by numerous cells. They
can mediate diverse biological functions, including antigen presentation. Exosomes have recently been shown to
contain functional RNA, which can be delivered to other cells. Exosomes may thus mediate biological functions
either by surface-to-surface interactions with cells, or by the delivery of functional RNA to cells. Our aim was
therefore to determine the presence of RNA in exosomes from human saliva, plasma and breast milk and whether
these exosomes can be taken up by macrophages.
Method: Exosomes were purified from human saliva, plasma and breast milk using ultracentrifugation and filtration
steps. Exosomes were detected by electron microscopy and examined by flow cytometry. Flow cytometry was
performed by capturing the exosomes on anti-MHC class II coated beads, and further stain with anti-CD9, anti-
CD63 or anti-CD81. Breast milk exosomes were further analysed for the presence of Hsc70, CD81 and calnexin by
Western blot. Total RNA was detected with a Bioanalyzer and mRNA was identified by the synthesis of cDNA using
an oligo (dT) primer and analysed with a Bioanalyzer. The uptake of PKH67-labelled saliva and breast milk
exosomes by macrophages was examined by measuring fluorescence using flow cytometry and fluorescence
microscopy.
Results: RNA was detected in exosomes from all three body fluids. A portion of the detected RNA in plasma
exosomes was characterised as mRNA. Our result extends the characterisation of exosomes in healthy humans and
confirms the presence of RNA in human saliva and plasma exosomes and reports for the first time the presence of
RNA in breast milk exosomes. Our results also show that the saliva and breast milk exosomes can be taken up by
human macrophages.
Conclusions: Exosomes in saliva, plasma and breast milk all contain RNA, confirming previous findings that
exosomes from several sources contain RNA. Furthermore, exosomes are readily taken up by macrophages,
supporting the notion that exosomal RNA can be shuttled between cells
Cupping is a therapeutic technique employed by many acupuncturists to treat Full,
Mixed or Deficiency conditions. Despite its documented effectiveness, however, little
is known from a Western scientific standpoint about the mechanisms by which
cupping therapy yields its effects. In 2005, Ilkay Chirali published the results of a
small trial conducted on 6 patients with musculoskeletal complaints. In this study
patients were treated every two days over a two weekly period and blood samples
were taken and analysed before, during and after the treatment period. Analysis of the
samples revealed a significant reduction in the Erythrocyte Sedimentation Rate
(ESR), a broad marker of inflammation. The aim of the current study was to identify
inflammatory marker(s) whose plasma concentrations are altered during the cupping
therapy in the treatment of musculoskeletal disorders and which may account for
changes in the ESR. This will provide biochemical evidence for the effectiveness of
cupping therapy.
Methodology
14 subjects presenting with a range of chronic musculoskeletal complaints (diagnosed
as Empty, Mixed and Full conditions) were recruited into the study. Full informed
consent was obtained from each subject. A Traditional Diagnosis was then performed
on each subject and the cupping treatment strategy determined accordingly. Each
subject then received six cupping treatments at weekly intervals. Prior to the first
treatment and thereafter at weekly intervals, subjects completed a MYMOP
questionnaire in order to assess clinical outcome. Venous blood samples were also
obtained before treatment, after 3 treatments, after 6 treatments and six weeks post
treatment. The following analyses were performed on blood samples: full blood
count, haemoglobin concentration, erythrocyte sedimentation rate (ESR), serum
concentration of fibrinogen, C Reactive Protein (CRP), IgG and ferritin. Serum
samples were also analysed for the concentration of inflammatory cytokines (IL-1β,
IL-6, IL-10 and TNF-α). Data obtained was analysed by one way ANOVA with
matched values and Dunnett’s Multiple Comparisons Test using Instat2 software.
Results
9 females (range 23 – 52 years) and 5 males (range 37 – 62 years) were recruited into
the study. One female participant subsequently withdrew from the study at week 3.
Analysis of MYMOP questionnaires indicated that 95% patients reported
improvements in their symptoms as a result of treatment and, overall, this
improvement was found to significant.
Complete data sets from blood and serum analysis (before, during and after treatment)
were obtained for 11 subjects. The six week post treatment follow up was 54%.
Analysis of data revealed statistically significant increases in the platelet count (p =
0.0373) and lymphocyte count (p = 0.0001) and decreases in the serum concentration
of fibrinogen (p = 0.0008) and ferritin (p = 0.0024). No changes were measured in the
serum concentration of proinflammatory cytokines, with the exception of one patient
where slight increases above normal levels were seen in TNF-α, IL-6 and IL-10
concentrations. Interestingly there was no significant change in the ESR observed in
these subjects, however, only 3 subjects presented with an elevated ESR before
treatment.
Discussion & Conclusion
Ferritin and fibrinogen are inflammatory markers and their reduction during cupping
therapy is concomitant with a reduction in the inflammatory status of the patient.
Platelets and lymphocytes may increase as a result of local vascular damage from the
cupping therapy itself or from a reduction in the adherence of these cells to areas of
activated endothelium, associated with inflammation. With regard to the ESR, no
overall significant change was observed in this study. However, only 3 patients
presented with an elevated ESR prior to treatment. Furthermore, the ESR did decrease
during the course of treatment in these subjects. In conclusion, although the precise
mechanism by which cupping therapy exerts it therapeutic effects cannot be
determined from the present study, the results indicate that cupping therapy is
associated with significant changes in the levels of inflammatory cells and soluble
markers suggesting that this treatment can influence the inflammatory status of the
patient leading to improved clinical outcome
A number of chemicals have been shown to demonstrate neurotoxic effects either in human or laboratory animal studies.
This article aims at evaluating the impact of exposure to several chemicals including: organophosphate, organochlorine
pesticides, polychlorinated biphenyls (PCBs), mercury and lead on the neurodevelopment of children by reviewing the
most recent published literature, and answer the question whether any progress has been made in the epidemiology
of the neurodevelopment of children induced by exposure to those chemicals. The result of the presented studies show
that exposure to the above-mentioned chemicals may impair the neurodevelopment of children. Neonates exposed to
organophosphate pesticides demonstrated a higher proportion of abnormal reflexes, and young children had more attention
problems. Exposure to organochlorine pesticides in children was associated with alertness, quality of alert responsiveness,
cost of attention and other potential attention associated measures. The majority of studies indicate the negative impact
of lead exposure at the level <10 μg/dl or even <5 μg/dl on the neurodevelopment of children. The results of studies on
exposure to PCBs, mercury, and their effect on neurodevelopment are inconsistent. Some suggest that prenatal exposure
to PCBs and mercury is related to performance impairments, attention and concentration problems, while other do not
present any statistically significant association. The studies were mostly well designed, using prospective cohorts with
the exposure assessment based on the biomarker of exposure. Concerning the covariates and confounders affecting the
endpoints in most of the presented studies, confounders were included in data analysis. In order to recognize the early
cognitive, motor and language outcomes of chemical exposures, well standardized tools were used for evaluating the
neurodevelopmental effects and offer an early and fairly comprehensive measure of child development. Because the
neurotoxicants may cross the placenta and the fetal brain, exposure consideration regarding the reduction of exposure to
those chemicals should be implemented.
Key words
pesticides, PCB, mercury, lead, neurodevelopment, exposure assessment