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Published in final edited form as:
. 2017 October ; 11(40): 621–634.
Revisiting the linkage between ethnomedical use and
development of new medicines: A novel plant collection strategy
towards the discovery of anticancer agents
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Joshua M. Henkin1, Kongmany Sydara2, Mouachanh Xayvue2, Onevilay Souliya2, A.
Douglas Kinghorn3, Joanna E. Burdette1, Wei-Lun Chen1, Bethany G. Elkington4, and Djaja
D. Soejarto1,*
1Department
of Medicinal Chemistry and Pharmacognosy, University of Illinois at Chicago, 833 S.
Wood St., Chicago, Illinois 60612, USA
2Institute
of Traditional Medicine, Ministry of Health, Vientiane Capital, Lao People’s Democratic
Republic
3Division
of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, Ohio State
University, 500 W. 12th Ave., Columbus, OH 43210, USA
4Science
and Education, Field Museum, 1400 S. Lake Shore Drive, Chicago, Illinois 60605, USA
Abstract
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The Vietnam-Laos International Cooperative Biodiversity Group (ICBG) based at the University
of Illinois at Chicago (UIC) catalyzed a country-wide network of medicinal plant preserves (MPP)
and medicinal biodiversity preserves (MBP) now established in ten provinces of the Lao People’s
Democratic Republic (Lao PDR), which are relied upon as protected sources of ethnomedicines
for local villagers and traditional healers. In collaboration with the Lao PDR’s Institute of
Traditional Medicine (ITM), our ongoing P01 Program Project (Ohio State University) examined
the anticancer bioprospecting potential for two of the most exhaustively inventoried of these sites:
the Bolikhamxay MPP and the Xiengkhouang MBP. Guided by prior voucher specimens sourced
from these preserves with an overwhelming emphasis on plants employed in traditional medicine,
201 distinct samples from 96 species were collected along with proper herbarium documentation.
Aliquots of these plant samples were extracted in azeotropic ethanol and evaporated to dryness for
initial biological evaluation. In six samples from six different species (2.99% of the collected
samples, 6.25% of taxa) it was observed that extracts exhibited notable cytotoxicity against HT-29
colon adenocarcinoma cells. The wisdom behind the utilization of HT-29 cells in this preliminary
biological screen is discussed. Furthermore, comparison of screening results based on
longstanding considerations and ideological underpinnings of ethnobotanical vs. “random”
biodiversity-based collection approaches is detailed herein. The results of this interdisciplinary
study support the hypothesis that, by privileging the initial sample set in terms of human safety
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*
Corresponding author: D.D. Soejarto. doelsoejarto@gmail.com.
Conflict of interest
The authors have not declared any conflict of interest.
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and pharmacological activity, ethnobotanically driven collection for biological screening efforts
can produce leads unprecedented by the strict traditional usages of plants.
Keywords
Lao PDR; medicinal plants; traditional medicine; cancer
INTRODUCTION
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In trusting human agency and the intentionality of traditional ecological knowledge, one
may be liable to conclude that botanical ethnopharmacopeias consist of plants that (A) bear
useful pharmacological activities and can improve health status and (B) can be administered
to patients in ways that render them clinically safe within reason (Fadeyi et al., 2013; Pan et
al., 2013; Getasetegn and Teferi, 2016). Given these two likelihoods, these
ethnopharmacopeias can be studied in their entirety through biological screening efforts
(Mazzio and Soliman, 2009; Fadeyi et al., 2013; Leonti and Weckerle, 2015; Odonne et al.,
2017), setting the stage in the drug discovery pipeline towards human health applications not
necessarily anticipated by indigenous knowledge and folk clinical application of the
particular plants evaluated in this way. It might even be expected that this strategy could
yield results superior to “random” collection efforts constrained to geographic areas with
similar levels of species richness and biodiversity (Bletter, 2007; Saslis-Lagoudakis et al.,
2012). Prior meta-analysis suggests that on a per sample basis, depending on ethnobotanical
use and screening assays performed for samples evaluated from Laos and Vietnam, plants
employed in traditional medicine can have a higher hit rate for bioactivity in empirical
studies (Gyllenhaal et al., 2012). Although there are caveats and nuances to this finding,
which will be discussed below, this insight is worth bearing in mind in the context of the
present paper. Our hypothesis accordingly states that the agentive, purposive nature of
botanical ethnopharmacopeias biases sample sets derived from them to useful bioactivity,
and thereby this selection criterion lends itself to success in initial biological screening and
drug discovery studies.
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Towards the end of further, and unambiguously, exploring the possibility for
ethnobotanically driven screening efforts, in this instance for the preliminary stages of
anticancer drug discovery, two of the most extensively inventoried preserves in the Lao
PDR, the Xiengkhouang MBP and the Bolikhamxay MPP, were selected as promising
expedition sites for our extramurally funded Program Project (P01) from among the ten
preserves that are extant (Sydara et al., 2014; Soejarto et al., 2015) (Figure 1). These
reservoirs for local traditional medicine plants served as the premiere P01 project sites for
the exploration of the ethnopharmacopeias of Laos through the lens of this pragmatic,
serendipitous-activity-through-human-utility paradigm. This paper presents the results of
this endeavor.
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METHODOLOGY
Memorandum of Agreement
A Memorandum of Agreement (MOA) for the conduct of collaborative research targeted to
the flowering plants, between the University of Illinois at Chicago and the Institute of
Traditional Medicine (ITM, Vientiane, Lao PDR), covering issues on intellectual property
and the sharing of benefits in the event of the discovery and development of a
pharmaceutical product was established. This MOA allowed for the collection of plant
materials (plant samples and their voucher herbarium specimens) in Laos and their transfer
to and biological evaluation in the USA.
Plant collection
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Following the signing of this agreement, a joint plant collection expedition between the
University of Illinois at Chicago (UIC) and the Institute of Traditional Medicine (ITM) was
undertaken in the Lao PDR. One expedition site was the Xiengkhouang Medicinal
Biodiversity Preserve (MBP) of the Kham District, Xiengkhouang Province, and the other
was Bolikhamxay Medicinal Plant Preserve (MPP) of the Paksan District, Bolikhamxay
Province (Figure 1).
Xiengkhouang Medicinal Biodiversity Preserve expedition
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The Xiengkhouang Medicinal Biodiversity Preserve (MBP) (as depicted in Figure 1) is
located about 50 km northeast of the capital city (Phonsavanh) of Xiengkhouang province,
near Ban Tha, a rural Lao Lum (Lowland Lao) village in the Kham District, and about 15
km from both Muang Kham and Ban Tha. The elevation is approximately 1140 m above sea
level, at 19°43′ N; 103°35′ E (GPS reading). This preserve comprises approximately 500
hectares of high quality, secondary, montane tropical rainforest (Figure 2).
Two of the ITM’s 5-passenger Toyota Hilux pickup trucks, with collapsible soft tops for
covering the rear cargo area, provided excellent mobility throughout the expedition period
(December 8–13, 2015). These vehicles allowed for the transportation of 6–10 passengers
(including drivers) in addition to the loads of collected plant materials and field supplies.
Two workers and several other locals were employed from the village of Ban Tha for the
purposes of harvesting and processing plant material.
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Two of the ITM’s 5-passenger Toyota Hilux pickup trucks, with collapsible soft tops for
covering the rear cargo area, provided excellent mobility throughout the expedition period
(December 8–13, 2015). These vehicles allowed for the transportation of 6–10 passengers
(including drivers) in addition to the loads of collected plant materials and field supplies.
Two workers and several other locals were employed from the village of Ban Tha for the
purposes of harvesting and processing plant material.
Near the Xiengkhouang MBP, quarters at the Seng Deuane guesthouse in Meuang Kham
were rented as the expedition base and as equipment and supplies storage. The concrete
patio of the guesthouse served as our base for drying plant samples and working quarters.
The space and facilities of this guesthouse permitted the efficient performance and
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completion of the expedition within a short time period. The warm and mostly dry
conditions in the winter season during the expedition eased the initial stages of processing of
the voucher specimens and the plant samples collected.
Part of the supplies and equipment for use in the expedition, such as tarpaulins, rice sacks,
nylon mesh bags, cardboard, plant presses, strings, branch cutters, twig clippers, knives
(pointed knives known locally as mid [/miː d/]), a GPS, digital cameras, binoculars, used
newspapers, and other plant collecting and processing supplies, was brought from Vientiane
through the ITM’s supplies or as part of the ITM and UIC personnel’s belongings.
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Forays into the field and the search for the plants to collect were guided by the list and
photographs of the plants previously collected (Soejarto et al., 2015) from this MBP. Plant
samples and voucher herbarium specimens were collected and carefully numbered and
documented by field notes and photographic images. Numbered herbarium specimens were
pressed between newspapers in the field. Screening samples, 1–3 kg fresh weight, depending
on nature or fleshiness of the plant material, were packed in sample collection bags made of
nylon mesh and were placed in the trucks. On return from the field, these were arranged in
rows to dry on the concrete patio of the Seng Deuane guesthouse, only being removed to the
interior of the guesthouse or a nearby wooden, roofed platform when the conditions were
overcast. At the end of the field operation (December 13, 2015), semi-dry samples in the
nylon mesh bags were loaded into the trucks, while voucher herbarium specimens in
newspapers were cinched in straps and cardboard sheets, and also loaded into the trucks. All
field equipment and plant materials collected were transported by the pickup trucks to the
ITM, where the drying of the samples and voucher herbarium specimens was completed.
Bolikhamxay Medicinal Plant Preserve expedition
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The Bolikhamxay Medicinal Plant Preserve (MPP), also known as the Somsavath MPP due
to its propinquity, usefulness, and political connection to Somsavath Village, is about 27 km
south of Pakxan, the capital city of Bolikhamxay province and is 163 m above sea level, at
18°27′ N; 103°48′ E. The preserve comprises approximately 13 hectares of high quality,
secondary, lowland broad-leaved tropical rainforests recovering from past fires and logging
(Figure 3), with adjoining land cleared for agriculture and plantations, primarily, economic
botanicals (Hevea rubber, agarwood, etc.).
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One of the ITM’s 5-passenger Toyota Hilux pickup trucks, with a collapsible soft top for
covering the rear cargo area, provided excellent mobility throughout the expedition period
(December 14–17, 2015). This vehicle allowed for the transportation of 3–5 passengers
(including drivers) in addition to the loads of collected plant materials and field supplies.
Guesthouse quarters in Pakxan were rented as the expedition base and as equipment and
supplies storage. The concrete patio and blacktop of the guesthouse served as our base for
drying plant samples and working quarters. The space and facilities of this guesthouse
permitted the efficient performance and completion of the expedition within a short time
period.
As in the Xiengkhouang expedition, part of the supplies and equipment for use in the
expedition was brought from Vientiane through the ITM’s supplies or as part of the ITM and
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UIC personnel’s belongings. In the collection area, one worker and several other locals were
employed from the village of Ban Khampai for the purposes of harvesting and processing
plant material. Forays into the field and the search for plants to be collected were guided by
a list and by photographs of the plants previously collected (Soejarto et al., 2015) from this
MPP. The warm and mostly dry conditions during the winter season of the expedition eased
the initial stages of processing for voucher specimens and screening samples in the field.
The roofed cement floor of the visitor center, near where the automobiles were usually
parked, served as a temporary processing and drying area before the plant materials were
loaded into the cargo bay of the Toyota Hilux on returning to the Pakxan guesthouse.
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Plant samples and voucher herbarium specimens were carefully numbered and documented
by field notes and photographic images. Numbered herbarium specimens were pressed
between newspapers in the field. Screening samples, 1–3 kg fresh weight, depending on
nature or fleshiness of the plant material, were packed in sample collection bags made of
nylon mesh and were placed in the truck during forays. Later these were arranged in rows on
the concrete patio of the guesthouse. At the end of the field operation (December 17, 2015),
semi-dry samples in the nylon mesh bags were loaded into the truck, while voucher
herbarium specimens in newspapers were cinched in straps and cardboard sheets. All were
transported by the pickup truck to the ITM, where the drying of the samples and voucher
herbarium specimens was completed.
Plant identification
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One set of voucher herbarium specimens was shipped to and processed and accessioned at
the John G. Searle Herbarium of the Field Museum of Natural History (F), Chicago. Two
additional sets of vouchers were deposited at the herbaria of the ITM. Taxonomic
identification of the plants collected was initially performed (by JMH, KS, OS, MX, DDS)
at the ITM Herbarium and was completed at the Herbarium of the Field Museum.
Plant extraction
Azeotropic ethanolic extracts of the plant samples were generated from 30 g aliquots of
plant samples at the laboratories of the ITM. Hence, 30 g aliquots of the 201 unground plant
samples were milled and subsequently macerated twice overnight in 250 mL, and then 200
mL of solvent, successively. The pooled extracts from each sample were desiccated by
rotary evaporation at 40 °C and transferred to small vials for shipment. All extracts were
dispatched to and received safely at the University of Illinois at Chicago, where they were
submitted to the MTS assay to determine their effect on the viability of HT-29 colon
adenocarcinoma cells.
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Colorimetric MTS assay for cell viability
Human colon cancer cells HT-29 were purchased from the American Type Culture
Collection (Manassas, VA). The cell line was propagated at 37°C in 5% CO2 in RPMI 1640
medium, supplemented with fetal bovine serum (10%), penicillin (100 units/ml), and
streptomycin (100 μg/ml). Cells in log phase growth were harvested by trypsinization
followed by two washings to remove all traces of the enzyme. A total of 5,000 cells were
seeded per well of a 96-well clear, flat-bottom plate (Microtest 96®, Falcon) and incubated
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overnight (37°C in 5% CO2). Samples dissolved in DMSO were then diluted and added to
the appropriate wells (concentrations: 20 μg/mL and 2 μg/mL; total volume: 100 μL;
DMSO: 0.5%). The cells were incubated in the presence of test substance for 72 hours at
37°C and evaluated for viability with a commercial absorbance assay (CellTiter 96®
AQueous One Solution Cell Proliferation Assay, Promega Corp Promega) that measured
viable cells. Survival percentage, based on microplate reader (Synergy Mx, BioTek) readings
of absorbance at 490 nm, was expressed in percentage relative to the solvent (DMSO)
control. One of the authors (Dr. Wei-Lun Chen) performed the bioassay and interpreted the
results (Ren et al., 2017).
RESULTS
Fieldwork
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A total of 201 plant samples for preliminary biological screening, comprising 96 species of
seed plants, documented by 96 sets of voucher herbarium specimens, were gathered during
the two expeditions (Table 1).
MTS assay
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At least six samples out of the 201 collected presented with activity bearing enough interest
for further testing with HT-29 colon adenocarcinoma cells using the colorimetric MTS assay
for cell viability, i.e. cytotoxicity. At the 20 μg/mL incubation condition, less than 60% of
the HT-29 cells in these samples survived, and in five out of six of these samples, less than
50% of the HT-29 cells survived, indicating that the IC50 of these five extracts should be
under 20 μg/mL. Given that all six samples were from different taxa, this means that 2.99%
of the collected samples and 6.25% of taxa with samples that were screened in this way were
sufficiently cytotoxic to merit initial recollection for further studies.
The six samples of interest (Table 2 and Figure 4) are the following: the stem material of
Cryptolepis dubia (Burm.f.) M.R.Almeida (A07194/ST; Asclepiadaceae); the aerial parts of
Rubia argyi (H.Lév. & Vaniot) Hara ex Lauener (A07196/PX; Rubiaceae); the fruits of
Reevesia pubescens Mast. (A07214/FR; Sterculiaceae); the combined leaves, twigs, and
fruits of Maclura tricuspidata Carrière (A07257/LF+TW+FR; Moraceae); the stem material
of Millettia pachyloba Drake (A07338/ST; Fabaceae-Papilionoideae); and the leaves and
twigs of Gardenia annamensis Pit. (A07365/LF+TW; Rubiaceae).
DISCUSSION
Usage of HT-29 and other human tumor cell lines in anticancer screening of plants
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The prevalence of cytotoxic taxa in this sample set derived from two expeditions in the Lao
PDR is generally consistent with the results anticipated by the laboratory personnel of the
P01CA125066 grant (Kinghorn et al., 2009; Kinghorn et al., 2016). The HT-29 cell line was
selected as the sole gatekeeper for initial cytotoxicity testing, from amongst a number of
human tumor cell lines accessible to Ohio State University scientists involved with the
program project. HT-29 was the only human tumor cell line on hand that displayed high
selectivity for the most cytotoxic extracts through its low susceptibility, with unpublished,
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internal project data suggesting that a signature of only ~5% (or less) of sample extracts
evaluated bore significant activity against HT-29 cells.
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In terms of the P01 project itself, the consequent focus on HT-29 cells and human colon
adenocarcinoma by association has led to screening for inhibitory activity in the K-ras
pathway becoming one of the major mechanistic assays emphasized (Peruchot et al., 1987;
Ren et al., 2014). HT-29 cells are also significant in that the cell line has long been and
continues to be included in the NCI-60 panel utilized for the COMPARE algorithm to
predict mechanism of action for cytotoxic compounds (Paull et al., 1989; Shoemaker, 2006).
More broadly HT-29 cells are derived from a human colon adenocarcinoma, and this subtype
of colon cancer, derived from the epithelial lining surrounding the lumen of the large
intestine, is responsible for over 90% of colon cancer cases (Kumar et al., 2010). According
to the Center for Disease Control’s most current data, furnished with the assistance of the
National Cancer Institute, as of 2013 colorectal cancer is the third most prevalent cancer
shared by both sexes in the United States of America and is the second deadliest (Center for
Disease Control, 2014).
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Aside from the P01 project’s established reasoning for proceeding first with cytotoxicity
screening using HT-29 cells under the aegis of the National Cancer Institute, the results of
this initial P01 expedition and bioassay work in the Lao PDR also show reasonable
consistency with prior bioprospecting findings of the Vietnam-Laos ICBG project (1998–
2012) (Gyllenhaal et al., 2012). It should be noted that the National Cooperative Drug
Discovery Group (“Novel Strategies for the Discovery of Plant-Derived Anticancer Agents”)
(Kinghorn et al., 2003; Balunas et al. 2006), the intellectual predecessor of the
P01CA125066 program project, overlaps substantially with the Vietnam-Laos ICBG
(Kinghorn et al. 2003; Kinghorn et al. 2009) in both the time period (1990–2006) and the
cell lines employed in cytotoxicity testing. Cell lines tested for viability against plant
extracts during the ICBG included COL-2 (human colon cancer); HL-60 (human
promyelocytic leukemia); hTERT-RPE1 (human telomerase reverse transcriptase-retinal
pigment epithelial cells); HUVEC (human umbilical vein endothelial cells); KB (human
cervical carcinoma, formerly believed to be oral carcinoma); LNCaP (human prostate
carcinoma); LU-1 (human lung cancer); and MCF-7 (human breast cancer) (Soejarto et al.,
2002; Kinghorn et al., 2003; Gyllenhaal et al., 2012; Zhang et al., 2016). Of these only
HUVEC and hTERT-RPE1 cells are not human tumor cell lines, and only HL-60 is clearly
not epithelial in origin. The NCDDG program utilized all of these cell lines as well as a
number of additional ones (Kinghorn et al., 2003). For the cell lines utilized in the ICBG
project, the cytotoxicity hit rate for Lao ethnomedical samples from plants ranged between
5% and 9%, with the exceptions of the non-epithelial leukemia cell line HL-60 (1.9%) and
the breast cancer cell line MCF-7, as no bioassays were performed with this latter cell line
for these samples (Gyllenhaal et al., 2012). No comparable meta-analysis in relation to the
cell lines utilized was produced by the NCDDG, although a synthesis communicated
significant insight into potential chemotaxonomic and plant anatomy-based relationships to
activity hit rates and levels of cytotoxicity (Balunas et al., 2006).
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Anticancer bioprospecting from plants: Medical ethnobotany collection, biodiversitybased collection, and the continuum in between
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Overall for the Lao ethnomedical plants on a per sample basis the hit rate for cytotoxicity in
cancer cells was higher whereas the “random” plants (from Vietnam) had a higher hit rate on
a per collection basis in the ICBG (Gyllenhaal et al., 2012). This disjunction can potentially
be explained in part through the fact that ethnomedical plant collection strategy often
samples only the plant part used locally whereas the “random” plant collection strategy
tends to sample as many plant parts per plant as can be managed, and therefore “random”
collection provides more chances for each taxon to possess a sample bearing activity in one
or more bioassays (Spjut, 2005; Gyllenhaal et al., 2012). For our expeditions outlined here
overall, for those taxa with collected samples, a little over two (2.09) samples per taxon were
obtained, some of which corresponded to plant parts used in local ethnomedicine and others
of which did not. Interestingly, 50% (three) of the active samples were from plant parts
employed from these taxa ethnomedically and the other 50% (three) were from plant parts
not used ethnomedically to our knowledge. For each plant that was active, only one sample
out of the 1–3 samples collected per taxon demonstrated significant cytotoxicity (Table 3).
None of these plants bearing active samples was employed locally to treat cancer. In this
context, these facts preliminarily suggest that the strategy of revisiting areas with
documented ethnopharmacopeias, collecting as many samples as possible per plant
harmonizes the benefits of “random” and ethnobotanical collection strategies for anticancer
screening while mitigating their respective downsides.
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Richard W. Spjut opines that while cytotoxicity to tumor cell lines and antitumor activity for
a plant taxon cannot be known a priori based on one reported ethnomedical application
versus another, comparing literature review and ethnopharmacological field work in tandem
with biological screening suggests in general that (A) greater toxicity categories exhibit
higher hit rates than all medicinal plants taken as one category and (B) certain categories had
three (anthelminthics) to four (arrow poisons and homicidal agents) times the hit rates of
plants screened from “random” collections (Spjut, 2005). Still it is not necessarily
straightforward that maximizing cytotoxicity in prioritizing bioactive leads always catalyzes
advancement in the US pharmaceutical pipeline, as evidenced by the development of antihepatitis C nucleoside analogues for instance, which was among the first preclinical
pharmacological research to suggest that it may behoove scientists to emphasize leads that
retain moderate activity while minimizing toxicity (Sluis-Cremer et al., 2009; Coats et al.,
2014).
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Plants and other sources of bioactive metabolites may be ideally positioned to take
advantage of the uncertain interplay between bioactivity and toxicity in the transition from
preclinical to clinical evaluation of leads, given that they often produce a suite of closely
related analogues. This is particularly salient for plants collected on the basis of use in local
medicine, by which their safety for human administration is comparatively and more likely
assured in contrast to other sources of bioactive compounds (combinatorial chemistry,
microbes, etc.). While as a result of his meta-analysis Spjut furthermore advocates
selectively mining medicinal plants for categorical, chemotaxonomic, and/or novelty-related
reasons in ongoing screening efforts, he also admits that the faster pace and greater sample
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collection rate for “random” (biodiversity-based) collection expeditions is attractive (Spjut,
2005). This is interesting because his analysis includes a further admission of the hybridity
of collection strategies (Spjut, 2005). This hybridity is bound up in multiple explanations as
to why the “random” collection strategy should be punctuated in quotations since, for
instance, phytogeography always and chemotaxonomy still often constrain the hit rate
results of plants selected for sampling from a given area.
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It should be noted that our Lao collection strategy for P01 anticancer screening resembles
the pace and sample collection rate of the “random” strategy that Spjut outlines far more
than the typical ethnobotany-driven or active sample re-collection expedition in his
experience, with an average of ~40/day obtained between the two expeditions, and within
the 1–3 active samples (out of 60–100 acquired) per day projected for a “random” collection
effort (Spjut, 2005). This also implies that at the very maximum, a “random” collection
effort might be expected to have twice the hit rate of these Lao P01 collection expeditions,
while at the low end, such “random” screening could have only a third of the hit rate we
observed. As with “random” collection expeditions for anticancer screening,
chemotaxonomic considerations had an impact on the samples we decided to collect (Spjut,
2005; Balunas et al. 2006). This principle along with related inherited wisdom from the
National Cancer Institute and the NCDDG/P01 project experience informed our exclusion of
certain taxa for sample collection throughout the expeditions. These built-in considerations
help demonstrate that this ethnobotanically driven screening strategy implemented for these
two Lao expeditions is not an example of ideological purity but rather of hybridity. We
would argue that this eclectic pragmatism contributes to the effectiveness of our innovative
strategy as a viable screening paradigm, which is supported perfectly by the fact that twice
the number of active hits was generated by broad collection of plant parts as would have
been through collection solely of plant parts employed in local ethnomedicine.
Prior results and integrative programing in Laos and Vietnam as a predictor of future
success and strategic considerations for medicinal plant bioprospecting
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The expansive and human-centric scope of the Vietnam-Laos ICBG has established and
funded infrastructure bolstering traditional medicine through the creation of rural preserves
and traditional medicine stations (TMS) in the Lao PDR (Riley, 2001; Sydara et al., 2014;
Soejarto et al., 2015); has permitted scholarly pursuit of botanical medicines described in
Lao palm leaf manuscripts (Elkington et al. 2009); and has thoroughly achieved the
inventory and preliminary drug discovery lead investigation of the medicinal plants, and the
floristic diversity as a whole, in these two countries (Soejarto et al., 2002; Soejarto et al.,
2006; Soejarto et al., 2012; Sydara et al., 2014). This project’s legacy is an exemplar of the
fact that medical ethnobotany, alongside such interdisciplinary subfields as historical
ecology, landscape archaeology, and political economy (Balée and Erickson, 2006;
Campbell, 2007; Scott, 2009), holds human agency and intentionality in high esteem, being
that they are crucial to value-added applications resulting from examination, discovery, and
rediscovery of traditional knowledge. Indeed, human-environment interactions and the
cultural transmission of natural and physiological observations are the bedrock underlying
the endeavors of medical ethnobotany and ethnopharmacology, ensuring their value in
perpetuity (Ott, 1998; Shepard, 2004)
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Consideration of past plant collection practices and biological screening results in Vietnam
and Laos to date should be able to guide the principles by which future expeditions are
performed in the collection of plant samples for the P01 project in the Lao PDR. For
instance, in the Vietnam-Laos ICBG, whereas cancer cell cytotoxicity hit rates for “random”
samples from Vietnam and ethnomedical samples from the Lao PDR were relatively high,
the ethnomedical samples from Vietnam had yielded a low hit rate (Gyllenhaal et al., 2012).
It has been suggested that the differences in these hit rates in ethnobotanical collections
between the two countries is attributable to the following factors: (a) a greater proportion of
accessions in Laos that were purported correctly by healers to treat cancer in particular and
(b) to a more specialized knowledge of medicinal properties of flora owing to the Lao
healers’ greater prominence and mastery (Gyllenhaal et al., 2012). While it has been
demonstrated that this strategy can lead to new bioactive leads for anticancer drug discovery,
category-focused ethnobotanical collection for biological screening seemingly is a tractable
and rewarding strategy for evaluating the medicinal potential of Lao plant biodiversity as
well.
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Although no bioprospecting and isolation work from the Lao PDR has thus far proceeded to
the stage of patent filing thereby meriting additional studies (unlike for Vietnam; Zhang et
al., 2014; Zhang et al., 2017), through the Vietnam-Laos ICBG, six medicinal plant taxa
from around the country have been evaluated through the compound isolation stage and have
been found to contain mostly novel phytochemicals bearing anti-tuberculosis (Elkington et
al., 2014); anti-malarial (He et al., 2005; He et al., 2006; Libman et al., 2008; Ma et al.,
2008); and cancer cell cytotoxicity activities (Zhang et al., 2004). With respect to anticancer
bioprospecting from the ICBG field work conducted from 1998–2012 in the Lao PDR, at
least 34 species yielded 50 extracts with significant activity (IC50 < 20 μg/mL) in one or
more cancer cell lines of a six cell line panel, which notably led to the isolated compounds
asparacoside and 3″-methoxy-nyasol (IC50 > 4 μg/mL, < 10 μg/mL) from Asparagus
cochinchinensis (Lour.) Merr. (Soejarto et al., 2012). Integrative research with plants
sourced from the network of ten preserves now established in Laos, to search for
unanticipated bioactivity, is only in the most incipient of stages. Given adequate resources,
biological screening and new active compound discovery from higher plants in these diverse
habitats of the Lao PDR could continue indefinitely for decades to come. The link between
bioprospecting results and the biodiversity of these preserves and the country overall is as
yet somewhat confounded by the underexplored relationship of abiotic and biotic factors to
the phytochemistry of higher plant taxa throughout the monsoon cycle and under local
management. These dynamic preserves are liable to yield additional surprises for as long as
interdisciplinary programs such as the P01 project are able to support additional expeditions
and follow-up research on taxa of interest (Kinghorn et al., 2009; Kinghorn et al., 2016).
CONCLUSIONS
The two initial P01 expeditions to the Lao PDR demonstrate that biological screening
efforts, particularly anticancer bioprospecting, can proceed in areas with ancient and
ongoing cultures of medicinal plant use, with the cooperation of local residents who protect,
manage, and depend on the various forest habitats of Laos. Given the unique opportunity for
direct collaboration with the Institute of Traditional Medicine (ITM) and some of the
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authors’ involvement with healers and protected areas throughout Laos, it is fully expected
that further expeditions will yield comparable or improved results. There are clear reasons
for why ethnobotanically motivated plant collection in areas rich in medicinal plant reliance
can pay greater dividends for drug discovery efforts than other strategies. These rationales
stem from the requirements of human safety, minimization of toxicity, and mitigation of
related side effect profiles, as well as efficacious pharmacological activity (as vetted by
longstanding use by local people), and from the pace of sample collection and turnover of
expedition results in terms of the substantial timetables for biological screening and
compound isolation work. The Bolikhamxay MPP and the Xiengkhouang MBP, which we
investigated during the winter dry season, are only two of the ten preserves established in the
country. It is hoped that more such preserves will be established in the future and maintained
locally to bolster resilience in medicinal plant resource use and management, ensuring longterm protection of these forested regions throughout Laos.
Author Manuscript
Far more inventory work and sample collection expeditions to these preserves would be
needed to truly exhaust the prospective bioactive leads from these ethnopharmacopeial
treasure troves of traditional plant medicine, even for anticancer screening alone. Our
interdisciplinary methodology and data analysis produce botanical and phytochemical leads
not documented by local plant use patterns, but on the whole – and verifiably – anticipated
by their value in ethnomedicine. The results of this research support our hypothesis that
investigating plants known to be employed in local ethnopharmacopeias can produce
promising starting points for natural product screening programs and pharmaceutical
development, particularly as a first stage for anticancer drug discovery.
Acknowledgments
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We wish to acknowledge the guidance of Dr. Yali Fu, who executed an SDCR status application for collectionsbased research of the National Institutes of Health/National Cancer Institute P01 project (P01CA125066) in the Lao
PDR. The status was awarded by the U.S. Department of State on September 9, 2015, which allowed for the
collection of voucher specimens and samples in Laos with the collaboration of the Institute of Traditional Medicine,
permitting material transfer to and biological evaluation in the USA. We wish to acknowledge the ethnobotany,
pharmacognosy, and chemistry staff of the Institute of Traditional Medicine for their involvement and support in the
project. This research could not have proceeded without the broad participation of the ITM staff, and the authors are
thankful for their tremendous effort and immense knowledge-base. We also wish to acknowledge the insights and
efforts of our workers on these two expeditions, notably Mr. Buasy in the Xiengkhouang MBP and Mr. Bounyong
in the Bolikhamxay MPP. Lastly, the participation of officials employed by the Food and Drug Divisions of the
Provincial Health Departments of Bolikhamxay and Xiengkhouang was essential for these expeditions to proceed,
and we are grateful to have solicited their enthusiastic involvement.
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Figure 1.
Medicinal Plants Preserve and Medicinal Biodiversity Preserves Network of Lao PDR
(Soejarto et al., 2015), showing the location of expedition sites described in this paper.
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Figure 2.
Xiengkhouang Medicinal Biodiversity Preserve, December 9–11, 2015. Top left: Roadside
signage downslope of the expedition site. Top right: View of the montane tropical rainforest
on a cloudy day with forbs and soil in the foreground. Bottom left: View of the tree cover
from the roadside. Bottom right: Forested rivulet within the preserve.
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Figure 3.
Bolikhamxay Medicinal Plant Preserve, December 15–16, 2015. Top left: Roadside signage
at the entrance of the preserve. Top right: View from the road of the path into the preserve,
on left. Bottom left: Processing samples on the patio of the pavilion near the entrance.
Bottom right: Lowland tropical rainforest within the preserve featuring trees, treelets, lianas,
and forbs.
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Figure 4.
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Plants from the Xiengkhouang Medicinal Biodiversity Preserve with samples exhibiting
notable cytotoxicity in HT-29 colon adenocarcinoma cells. A: Cryptolepis dubia. B: Rubia
argyi. C: Reevesia pubescens. D: Maclura tricuspidata. E: Millettia pachyloba. F: Gardenia
annamensis.
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Table 1
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Plant samples from the December 2015 P01 expeditions in the Lao PDR, listed in order of their voucher
herbarium collection and by their scientific name (species-family), number and part of the plant of the primary
screening samples, and locations. Plant part abbrevations: FL (flowers); FR (fruits); LF (leaves); PL (whole
plant); PX (aerial parts); RT (roots); SB (stem bark); ST (stem); SW (stem wood); TW (twigs).
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Voucher herbarium specimen
Species (Family)a
JMH 001
Medinilla septentrionalis
(W.W. Sm.) H.L. Li
(Melastomataceae)
A07186/ST; A07187/RT; A07188/LF+TW+FL+FR
JMH 003
Micromelum falcatum
(Lour.) Tanaka (Rutaceae)
A07189/RT; A07190/ST; A07191/LF+TW+FR
JMH 004
Derris scandens (Roxb.)
Benth. (FabaceaePapilionoideae)
JMH 005
Cryptolepis dubia (Burm.f.)
M.R. Almeida
(Asclepiadaceae)
JMH 006
Rubia argyi (H.Lév. & Van.)
Hara ex Lauener
(Rubiaceae)
JMH 008
Actinodaphne rehderiana
(C.K. Allen) Kosterm.
(Lauraceae)
A07197/ST; A07198/LF+TW
JMH 009
Cissampelos pareira L.
(Menispermaceae)
A07199/PX
JMH 010
Clematis leschenaultiana
DC. (Ranunculaceae)
A07200/PX
JMH 011
Vitex quinata (Lour.) F.N.
Williams (Verbenaceae)
A07201/FR; A07202/LF+TW; A07203/ST
JMH 012
Schefflera cf. leucantha R.
Vig. (Araliaceae)
A07204/FR+TW; A07205/LF+TW; A07206/ST
JMH 013
Elsholtzia blanda (Benth.)
Benth. (Lamiaceae)
A07207/PX
JMH 014
Saurauia napaulensis DC.
(Actinidiaceae)
A07208/FR+TW; A07209/LF+TW
JMH 016
Clematis subumbellata Kurz
(Ranunculaceae)
A07210/PX
JMH 017
Mucuna bracteata DC.
(Fabaceae-Papilionoideae)
A07211/PX
JMH 018
Ilex sp. (Aquifoliaceae)
A07212/ST; A07213/LF+TW+FR
JMH 021
Reevesia pubescens Mast.
(Sterculiaceae)
A07214/FR; A07215/LF+TW; A07216/ST
JMH 022
Schima wallichii (DC.)
Korth. (Theaceae)
A07217/LF+TW+FR; A07218/ST
JMH 023
Rourea minor (Gaertn.)
Alston (Connaraceae)
A07219/ST; A07220/LF+TW; A07221/FR
JMH 024
Acacia pennata (L.) Willd.
(Fabaceae-Mimosoideae)
A07222/FR; A07223/LF+TW; A07224/ST
JMH 025
Wendlandia uvariifolia
Hance ssp. laotica (Pit.)
Cowan (Rubiaceae)
Primary samples (Plant parts)
Collection location
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
A07192/ST; A07193/LF+TW
Xiengkhouang MBP
A07194/ST; A07195/LF+TW
Xiengkhouang MBP
A07196/PX
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
A07225/FR; A07226/LF+TW
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Voucher herbarium specimen
Species (Family)a
Primary samples (Plant parts)
JMH 026
Wikstroemia meyeniana
Warb. (Thymelaeaceae)
A07227/ST; A07228/RT; A07229/LF+TW+FR
JMH 027
Engelhardia roxburghiana
Wall. (Juglandaceae)
A07230/SB; A07231/ LF+TW+FR; A07232/SW
JMH 028
Rhus chinensis Mill.
(Anacardiaceae)
A07233/LF+TW+FR
JMH 029
Rotheca serrata (L.) Steane
& Mabb. (Verbenaceae)
A07234/LF+TW+FL; A07235/ST
JMH 030
Vernonia arborea Buch.Ham. (Asteraceae)
A07236/LF+FL; A07237/ST
JMH 031
Uncaria sessilifructus Roxb.
(Rubiaceae)
A07238/LF+TW+FL; A07239/ST
JMH 032
Debregeasia longifolia
(Burm.f.) Wedd.
(Urticaceae)
A07240/LF+TW+FR; A07241/ST
JMH 036
(Lamiaceae?)
A07242/PX
JMH 037
Casearia graveolens Dalzell
(Flacourtiaceae)
A07243/LF+TW+FR; A07244/ST
JMH 038
Pittosporum napaulense
(DC.) Rehder & E.H.
Wilson (Pittosporaceae)
A07245/LF+TW+FR; A07246/ST
JMH 039
Eurya laotica Gagnep.
(Theaceae)
A07247/LF+TW+FR; A07248/ST
JMH 040
Melastoma imbricatum
Wall. ex Triana
(Melastomataceae)
A07249/LF+TW; A07250/ST
JMH 041
Rubus alceifolius Poir.
(Rosaceae)
A07251/LF+TW; A07252/ST
JMH 042
Rubus pluribracteatus
L.T.Lu & Boufford
(Rosaceae)
A07253/LF+TW; A07254/ST
JMH 043
Ligustrum sinense Lour.
(Oleaceae)
A07255/LF+TW; A07256/ST
JMH 044
Maclura tricuspidata
Carrière (Moraceae)
A07257/LF+TW+FR; A07258/ST
JMH 045
Elephantopus mollis Kunth
(Asteraceae)
A07259/PL
JMH 046
Tadehagi triquetrum (L.) H.
Ohashi (FabaceaePapilionoideae)
A07260/PL
JMH 048
Itea macrophylla Wall.
(Iteaceae)
A07261/LF+TW+FR; A07262/SB; A07263/SW
JMH 049
Rhynchotechum ellipticum
(Wall. ex D. Dietr.) A. DC.
(Gesneriaceae)
A07264/LF+FR; A07265/ST
JMH 052
Symplocos lancifolia Sieb.
& Zucc. (Symplocaceae)
A07266/LF+TW; A07267/SB; A07268/SW
JMH 053
Chloranthus spicatus
(Thunb.) Makino
(Chloranthaceae)
A07269/PX
JMH 055
Buddleja asiatica Lour.
(Buddlejaceae)
A07270/LF+TW+FL; A07271/ST
Collection location
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
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Voucher herbarium specimen
Species (Family)a
Primary samples (Plant parts)
JMH 057
Cayratia tenuifolia (Wight
& Arn.) Gagnep. (Vitaceae)
A07272/LF+TW+FR; A07273/ST
JMH 058
Oreocnide integrifolia
(Gaudich.) Miq.
(Urticaceae)
A07274/LF+TW+FR; A07275/ST
JMH 059
Gynura divaricata (L.) DC.
(Asteraceae)
A07276/PX
JMH 060
Deeringia amaranthoides
(Lam.) Merr. (Rosaceae)
A07277/PX
JMH 061
Anneslea fragrans Wall.
(Theaceae)
A07278/LF+TW; A07279/SB
JMH 062
Blumea sp. (Asteraceae)
A07280/PX
JMH 063
Tithonia diversifolia
(Hemsl.) A.Gray
(Asteraceae)
A07281/LF+TW+FL; A07282/ST
JMH 065
Engelhardia spicata Lesch.
ex Bl. (Juglandaceae)
A07283/LF+TW; A07284/SB; A07285/SW
JMH 068
Omphalea bracteata
(Blanco) Merr.
(Euphorbiaceae)
A07286/ST; A07287/LF+TW
JMH 069
Baccaurea ramiflora Lour.
(Euphorbiaceae)
A07288/LF+TW; A07289/ST
JMH 070
Vitex stylosa Dop
(Verbenaceae)
A07290/LF+TW; A07291/ST
JMH 071
Ancistrocladus tectorius
(Lour.) Merr.
(Ancistrocladaceae)
JMH 072
Lasianthus trichophlebus
Hemsl. ex F.B. Forbes &
Hemsl. (Rubiaceae)
A07294/LF+TW; A07295/ST
JMH 073
Psychotria cephalophora
Merr. (Apocynaceae)
A07296/LF+TW; A07297/ST
JMH 074
Dracaena cambodiana
Pierre ex Gagnep.
(Agavaceae)
A07298/LF+TW; A07299/ST
JMH 075
Gardenia cf. annamensis Pit.
(Rubiaceae)
A07300/LF+TW; A07301/ST
JMH 076
Eurycoma longifolia Jack
(Simaroubaceae)
A07302/LF+TW; A07303/ST
JMH 077
Mussaenda glabra Vahl
(Rubiaceae)
A07304/LF+TW; A07305/ST
JMH 078
Bauhinia penicilliloba
Pierre ex Gagnep.
(FabaceaeCaesalpinioideae)
A07306/LF+TW; A07307/ST
JMH 079
Gnetum macrostachyum
Hook. f. (Gnetaceae)
A07308/PX
JMH 080
Breynia fleuryi Beille
(Euphorbiaceae)
A07309/PX
JMH 081
Garcinia celebica L.
(Clusiaceae)
A07310/LF+TW; A07311/ST
JMH 082
Connarus paniculatus Roxb.
(Connaraceae)
A07312/LF+TW; A07313/ST
Collection location
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Xiengkhouang MBP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
A07292/LF+TW; A07293/ST
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
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Voucher herbarium specimen
Species (Family)a
Primary samples (Plant parts)
JMH 083
Kibatalia laurifolia (Ridl.)
Woodson (Apocynaceae)
A07314/LF+TW; A07315/ST
JMH 084
Cratoxylum formosum
(Jacq.) Benth. & Hook. f. ex
Dyer (Clusiaceae)
JMH 085
Knema erratica (Hook. f. &
Thomson) J. Sinclair
(Myristicaceae)
A07317/LF+TW; A07318/SB; A07319/SW
JMH 086
Pterospermum argenteum
Tardieu (Sterculiaceae)
A07320/LF+TW; A07321/SB; A07322/SW
JMH 087
Arenga caudata (Lour.) H.E.
Moore (Arecaceae)
A07323/PX
JMH 088
Lithocarpus cf.
toumorangensis A. Camus
(Fagaceae)
JMH 089
Jasminum cf. annamense
Wernham ssp. glabrescens
P.S.Green (Oleaceae)
A07327/PX
JMH 090
Alpinia calcarata (Haw.)
Roscoe (Zingiberaceae)
A07328/LF; A07329/ST; A07330/RT
JMH 091
Amomum cf. villosum Lour.
(Zingiberaceae)
A07331/LF; A07332/ST; A07333/RT
JMH 092
Thysanolaena cf. latifolia
(Roxb. ex Hornem.) Honda
(Poaceae)
JMH 093
Cyperus trialatus
(Boeckeler) J. Kern
(Cyperaceae)
A07336/PL
JMH 094
Millettia pachyloba Drake
(Fabaceae-Papilionoideae)
A07337/LF+TW; A07338/ST
JMH 095
Saccharum arundinaceum
Retz. (Poaceae)
A07339/PX
JMH 096
Miscanthus cf. sinensis
Andersson (Poaceae)
A07340/LF; A07341/ST
JMH 097
Lagerstroemia balansae
Koehne (Lythraceae)
A07342/LF+TW; A07343/SB; A07344/SW
JMH 098
Barringtonia pauciflora
King (Lecythidaceaee)
A07345/LF+TW; A07346/ST
JMH 099
Ormosia cambodiana
Gagnep. (FabaceaePapilionoideae)
A07347/LF+TW; A07348/SB; A07349/SW
JMH 100
Aporosa ficifolia Baill.
(Euphorbiaceae)
A07350/LF+TW; A07351/ST
JMH 101
Ardisia conspersa E. Walker
(Myrsinaceae)
A07352/LF+TW+FR; A07353/ST
JMH 102
Securidaca inappendiculata
Hassk. (Polygalaceae)
A07354/LF+TW; A07355/ST
JMH 103
Syzygium cf. chloranthum
(Duthie) Merr. & L.M.Perry
(Myrtaceae)
A07356/LF+TW; A07357/ST
JMH 104
Peltophorum dasyrrhachis
(Miq.) Kurz (FabaceaeCaesalpinioideae)
A07358/LF+TW; A07359/SB; A07360/SW;
A07361/RT
Collection location
Bolikhamxay MPP
Bolikhamxay MPP
A07316/ST
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
A07324/LF+TW; A07325/SB; A07326/SW
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
A07334/LF; A07335/ST
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
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Voucher herbarium specimen
Species (Family)a
JMH 105
Capparis trinervia Hook. f.
& Thomson
(Capparidaceae)
A07362/ST
JMH 106
Aporosa tetrapleura Hance
(Euphorbiaceae)
A07363/LF+TW; A07364/ST
JMH 107
Gardenia annamensis Pit.
(Rubiaceae)
A07365/LF+TW; A07366/SB; A07367/SW
JMH 108
Macaranga denticulata (Bl.)
Muell.-Arg.
(Euphorbiaceae)
A07368/LF+TW; A07369/SB; A07370/SW
JMH 109
Maesa ramentacea (Roxb.)
A. DC. (Myrsinaceae)
A07371/LF+TW; A07372/ST
JMH 111
Sandoricum koetjape
(Burm.f.) Merr. (Meliaceae)
A07373/LF+TW; A07374/SB; A07375/SW
JMH 112
Tetrameles nudiflora R. Br.
(Tetramelaceae)
A07376/LF+TW; A07377/SB; A07378/SW;
A07379/RT
JMH 113
Adenanthera pavonina L.
(Fabaceae-Mimosoideae)
A07380/LF+TW; A07381/SB; A07382/SW
JMH 114
Fernandoa cf. adenophylla
(Wall. ex G. Don) Steenis
(Bignoniaceae)
A07383/LF+TW; A07384/ST
JMH 115
Uncaria sinensis (Oliv.)
Havil. (Rubiaceae)
A07385/LF+TW; A07386/ST
Primary samples (Plant parts)
Collection location
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
Bolikhamxay MPP
a
Traditional family names for each species listed are used in this paper; for current family names, please consult Tropicos (http://
www.tropicos.org/), The Plant List (http://www.theplantlist.org/) or APG III (http://www.mobot.org/MOBOT/research/APweb/).
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Table 2
Author Manuscript
Plants from the Xiengkhouang Medicinal Biodiversity Preserve with samples exhibiting notable cytotoxicity
in HT-29 colon adenocarcinoma cells.
Active Species
Active Sample
(Corresponding Voucher
Specimen)
Plant Part(s) Active
% HT-29 Cell
Survival (2 µg/mL)
% HT-29 Cell
Survival (20
µg/mL)
Cryptolepis dubia
A07194 (JMH 005)
Stem (ST)
100
48
Rubia argyi
A07196 (JMH 006)
Aerial parts (PX)
69
36
Reevesia pubescens
A07214 (JMH 021)
Fruits (FR)
84
24
Maclura tricuspidata
A07257 (JMH 044)
Leaves, twigs, and fruits (LF+TW
+FR)
79
56
Millettia pachyloba
A07338 (JMH 094)
Stem (ST)
77
33
Gardenia annamensis
A07365 (JMH 107)
Leaves and twigs (LF+TW)
100
37
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Table 3
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Comparison of plants and plant parts active with their local employment in traditional medicine.
Active Species
Plant Part(s) Active Against
HT-29 Cells
Lao Local Name
Plant Part(s) Used
in Traditional
Medicine
Traditional Use
Cryptolepis dubia
Stem (ST)
Kheua en one
Liana
Tonic for tendon and muscle
Rubia argyi
Aerial parts (PX)
Kheua lin ma nai
Whole liana
Fever, sore throat, kidney stone
Reevesia pubescens
Fruits (FR)
Mai sa fay
Root, stem
Gastritis
Maclura tricuspidata
Leaves, twigs, and fruits (LF+TW
+FR)
Kok nam thaeng
Root
Kidney edema (swollen kidney),
tonic for mother after giving birth
Millettia pachyloba
Stem (ST)
Xa kheuy done
Liana
Laxative
Gardenia annamensis
Leaves and twigs (LF+TW)
Khai nao
Stem
Stomachache
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