ASPECTS OF ANCIENT MITOCHONDRIAL
DNA ANALYSIS IN DIFFERENT POPULATIONS
FOR UNDERSTANDING HUMAN EVOLUTION
Nesheva DV
*Corresponding Author: Desislava V. Nesheva, Department of Medical Genetics, Medical Faculty, Medical University,
2 Zdrave str., fl. 6, 1431 Sofia, Bulgaria. Tel.: +35-92-917-2735; E-mail: desislava.nesheva@gmail.com
page: 5
|
|
INTRODUCTION
of the nucleus in cytoplasm of the cells. They
play a central role in cell life and death. They have
small genome, independent of the nuclear DNA-mitochondrial
DNA (mtDNA). Mitochondrial DNAs
are circular, double-stranded molecules, with high
copy number, and a higher evolutionary importance
compared to nuclear DNA. They have specific uniparental
inheritance only from mothers to their child,
which is useful for tracing matrilineal kinship in
many generations [1-4].
Mitochondrial DNA is a proper tool for determination
of the origin of different populations. It is
an important object of study in different fields such
as evolutionary anthropology, population genetics,
medical genetics, genetic genealogy and forensic
science [1,5,6].
Mitochondrial DNAs are not involved in recombinant
processes and their variants are due only
to mutations. Mitochondrial DNA is characterized
by a high mutation rate that allowed the sequential
accumulation of neutral mutation-specific base replacements,
especially transitions, which arose approximately
at the same time, when people inhabited
different regions all over the world. These mutations
form groups of stable haplotypes and are known as
haplogroups. Mitochondrial DNA haplogroups tend
to be geographically restricted and they are used to
genetically distinguish populations [5,6].
Mitochondrial DNA Variability. The most
variable part of mtDNA is the control region [displacement loop (D-loop)].It is the biggest non coding
part of mtDNA and plays a role in regulation and
initiation of replication and transcription. The most
polymorphic sequences in the control region are the
hypervariable segment I and hypervariable segment
II (HVS I and HVS II, respectively). They are objects
of many studies and researches of the roots of
populations and human evolution [7,8].
The mtDNA variation data can be used for creating
genealogical trees that contain information about
the order of the evolutionary processes in space and
time. The first mtDNA tree was created by Vigilant
et al. [9], and the first five branching points of the
mtDNA tree were from people living in sub-Saharan
Africa. The understanding of the evolution of the
mtDNA pedigree helped population geneticists to
trace the ancestors of modern humans to their roots in
Africa and their subsequent distribution in the world.
It was determined that the Mitochondrial Eve lived in
sub-Saharan Africa 200,000 years ago. As shown by
fossils found in Israel, the earliest human invasions
out of Africa started from the Kalahari Desert and the
African rainforest 90,000 years ago. The successful
migratory processes out of Africa has been proven by
mtDNA data and is dated to 55,000 to 85,000 years
ago. Researchers created the theoretical roads of migrations
along the Nile and across the Sinai Peninsula
spreading all over the world. Obtained data show that
Australia and Asia were first to be inhabited [10],
whereas Europe was initially colonized 45,000 years
ago. Other important events in European prehistory
are from 27,000 to 16,000 years ago at the Last Glacial
Maximum forming the uninhabitable areas in
Europe and between 9000 and 5000 years ago with
the spread of the Neolithic culture in Europe [11].
Mitochondrial DNA Haplogroups. Today, the
oldest mtDNA haplogroups are found in Africa. The
first haplo-groups were L1, L2 and L3, and they gave
rise to other macro-haplogroups and branches of the
global phylogenetic tree during the migration waves
from Africa all over the world. Haplogroup L3 is ancestral
to macro-haplo-groups M and N. They arose
in northeast Africa and spread into Europe and Asia.
Haplogroups H, I, J, N1b, T, U, V, W and X derived
from haplogroup N, and at present they comprise
the majority of mtDNAs in Europe. The Asian haplogroups
A, B, C, D, F and G derived from M and
N. Haplogroups A, B, C and D are frequent among
Native Americans [1].
Mitochondrial Gene Pool. The European mtDNA
gene pool is quite homogeneous. The comparison
of the sequences from Europe and other continents
revealed that the genetic distances between European
populations are much lower than between the
populations from other continents. In the phylogenetic
tree of several world populations, the European
population forms a small cluster near Turkey and the
Middle East. The formation of the modern European
gene pool is probably complex, slow and a relatively
recent process [12]. Recent studies show that over
the past few millennia, populations living in northeastern
Europe and Asia had close contacts. This is
evidenced by different migration waves that crossed
the Bosporus in both directions. The formation of the
European populations was largely influenced by two
large population expansions from the Middle East to
the West (the initial colonization of the continent and
the Neolithic expansion) [13].
The human mtDNA variations are being recorded
by aligning mtDNA sequences to the revised
Cambridge reference sequence (rCRS). The rCRS
is the corrected version of the first fully sequenced
mtDNA genome and belongs to haplogroup H. The
CRS was created in 1981 and revised in 1999 [14].
For the first time mtDNA was sequenced during the
1970s from a group under Dr. Fred Sanger at Cambridge
University, Cambridge, Cambridgeshire, UK.
Ancient DNA. The development and improvement
of technologies allowed the retrieval of DNA
sequences from museum specimens, archaeological
finds and fossil remains. This is the initial basis of
ancient DNA (aDNA) research. Analyses and comparisons
of ancient and modern mtDNA can provide
evidence for the population origin and migration
processes [15,16]. In 1983, Higuchi et al. [17]
at Berkeley, CA, USA, first successfully extracted
and sequenced ancient mtDNA from dried muscle,
a 150-year-old museum specimen of the quagga, a
zebra-like species. In 1985, Paabo studied Egyptian
mummified material, the first human remains dating
as far back as several thousand years [18]. In 1997,
aDNA from Neanderthal specimens excavated from
the Feldhofer Cave in Germany was successfully
extracted [19].
Despite the doubt about the success of the investigations
in the dawn of aDNA studies, the first
results gave new directions in science such as the
development of anthropogenetics and paleogenetics and increased their role in the evolutionary science.
In paleogenetic analyses, scientists sequenced
the hypervariable fragments of mtDNA extracted
from skeletal remains recovered from archaeological
sites, they compared the results with literature
data and classified mtDNAs into maternal lines according
to the sequence polymorphism. This was a
very useful approach and enabled the identification
of the genealogy of individuals with a high level of
authenticity. The genetic structure, origins, changes,
migration routes of ancient people could certainly be
more clearly understood. Most of the examinations
for tracing ancient human movements and obtaining
dates for genetic prehistory are now routinely
performed. Their main idea is to create chronology
linking living contemporary humans with their ancestors.
The examinations of ancient populations in
the last 10 years strongly imply the major role of
ancient mtDNA studies in the determination of features
of the prehistoric migrations and demographic
expansions [20,21].
Authenticity and Contamination. The main
problem with authenticity of aDNA is contamination
with exogenous, mostly contemporary DNA;
postmortem damages; errors during work in the
multiple independent amplification and sequencing
processes; degradation and low copy number of
partly preserved mtDNA. These problems are sufficient
reason to create criteria for working with old
samples. Such criteria were suggested by Cooper and
Poinar [22]. These criteria are useful for establishing
the methods and techniques for working with
these kinds of samples. The implementation of these
strategies will help to have effective and successful
results from ancient mtDNA [22]. On the other
hand, the Nine Criteria will elevate the chances for
authentic results in ancient mtDNA extraction and
sequencing. The Nine Criteria include repeatability
of experiments from two separate extracts, necessary
to prove previous results. Posterior controls of
the samples are performed to avoid the presence of
traces or sequences that can be from the excavation
team, curating staff, or laboratory personnel rather
than mtDNA lineages that would be expected. In
other words, posterior controls are searching for the
presence of mosaic or abnormal structure that can be
evidence of contamination. When the combination
from different sequenced fragments of aDNA are not
close to any point in the phylogeny or have an unusual
combinations of mutations, they are considered
to be a result of postmortem changes and phantom
mutations. The scientists noted that checking work
at every step will help to provide and confirm the
authenticity of aDNA [23,24]. Another important
point is working in a high level of sterile conditions.
The monitoring of contamination should start during
the excavation of the remains. The results from the
sequencing of the samples should be compared with
the sequences of all the people who are performing
the analyses. The ancient and modern DNA laboratories
should be separate, and the working surfaces
should be UV irradiated [25]. Researchers planning
to perform aDNA studies have to be able to follow
all necessary criteria for aDNA work.
There is a theory for the discrimination of contaminant
from endogenous sequences of mtDNA,
according to which contaminant sequences are longer
than the endogenous one, because in ancient samples
DNA is degraded from long time preservation. The
length of the obtained sequences can be directly observed
using shotgun sequencing of mtDNA. On the
basis of this theory, the scientists found that up to
80.0% of the longest sequences of the Neanderthal
were modern human contaminations. Further analyses
have shown that endogenous and contaminant
sequences presented different length distributions
and the size reduction depends upon different factors
such as age of the samples and level of contamination.
The theory is not useful in some cases and especially
in a particular set of sequences because there is
wide overlapping of the length distributions in different
samples. But even this, the scientists concluded
that these criteria can be applied for measuring the
authenticity with increased confidence for all ancient
high-throughput sequencing data sets [25,26].
The retrieval of mtDNA from ancient human
specimens is not always successful owing to DNA
deterioration and contamination. Usually only short
DNA fragments can be retrieved from these ancient
specimens. Even though ancient DNA provided the
limited amount of data in the past, it is the main
resource for direct information about our ancient ancestors.
The analyses of ancient samples can provide
new perspectives on human history, but the problem
with degradation and contamination in long-term
preserved specimens still makes analyses very difficult.
This is due to the technical difficulties with
extraction, amplification and sequencing of ancient mtDNA. The scientists tried to improve different
methods and techniques for working with ancient
mtDNA, such as polymerase chain reaction (PCR)
with specifically constructed primers for short overlapping
fragments covering large-size sequences
containing haplogroup-diagnostic mutations, cloning
and sequencing. There are still no absolutely perfect
techniques for obtaining authentic aDNA, but researchers
are working on finding them [15].
Materials and Methods for Ancient Mitochondrial
Phylogenetic Analyses. The main materials
that can be used for extracting ancient mtDNA
are usually dry hard remains: roots of teeth or pieces
of femur. They are the appropriate source of aDNA,
because they preserve DNA for a long period of time.
There are several well established protocols for extracting,
amplification and sequencing of ancient
mtDNA (especially HVS I and HVS II). Extraction
of mtDNA from bone powders usually follows the
protocols and controls for sterility, as phenol-chloroform
or silica-based protocols are usually applied
[27,28]. Several steps of amplification of fragments
of mtDNA are necessary because DNA is fragmented
due to long-term preservation. The fragments are
overlapped, which allows the covering of the whole
mtDNA genome or the HVS I and HVS II regions. In
the next step of the protocol, the fragments are cloned
by using specific competent cells (E. coli) that allows
control of the fragments’ quality and the screening
of the correctly inserted clones [29]. The Sanger et
al. [30] method is primarily used for the sequencing.
In the last 5 years, there have been attempts to
apply next generation sequencing as a method for
analysis of ancient samples. These attempts show
that next generation sequencing is suitable for aDNA
research [31].
Neanderthals and Anatomically Modern Humans.
Ancient DNA studies demonstrated their potential
to help us to understand the macro-evolutionary
history, particularly when it came to clarify the
relationships between existing and extinct species.
Working with ancient mtDNA can prove or reject the
theory that Neanderthals and anatomically modern
humans contributed in varying degrees to the modern
human gene pool. Some studies of Nean-derthals and
early humans show that they did not intermingle.
Other studies show different results. Thus, the sequencing
of HVRI of 23,000- and 25,000-year-old
bones from ancient people reveal that they are not so
different from contemporary people but they differ
from Nean-derthals. This conception is in the basis of
the multire-gional hypothesis of modern human origins
[32]. There are studies showing that the evolutionary
history of Neander-thals and modern humans
is characterized by similar demographics parameters
[33,34]. These surveys try to prove the hypothesis for
interbreeding between Neanderthals and anatomically
modern humans. They found changes in the
morphology of late Neanderthals and observed some
features in anatomically modern humans that are influenced
from Neanderthals. The examination of the
mandible from Riparo Mezzena, a Middle Paleolithic
rock shelter in the Monti Lessini, Verona, Italy,
showed that it was anatomically typical of modern
humans but genetically and morphologically typical
of late Neanderthals [35-37]. The study confirms that,
this change in the morphology of the mandibular chin
of Mezzena and other late Neanderthals, could be the
result of a small degree of interbreeding with anatomically
modern humans or genetic admixtures between
Homo sapiens and Homo neanderthalensis [35,38].
This examination will help for better understanding
the transition between these to human groups. The
changes in the morphology in late Neanderthals lend
support to the hypothesis of continuity with anatomically
modern humans or interbreeding with them.
Changes During the Neolithic and Formation
of the European Gene Pool. In a study of archaeological
and paleoecological data of ancient samples
(in 1995) it was shown that genetic differences observed
in Basques are compatible with migration during
the last glacial period around 18,000 years. After
the spread of modern humans in Europe, high levels
of migration, particularly in the Neolithic period,
probably contributed to the homogenization of the
gene pool in Europe [39,40]. About 11,000 years ago
there was replacement of the hunting-and-gathering
lifestyle in the Near East with agriculture. Agriculture
spread into Europe about 7500 years ago and it
reached from Hungary and Slovakia to Ukraine in the
east and to France in the west. There is an on-going
archeological, anthropological and population genetic
debate whether the spread of farming involved
large farmers migrations, i.e., demic diffusion or if
it was due to cultural diffusion. A study comparing
ancient DNA from the early farmers and hunter-gatherers
shows that farmers were genetically so distinct
from hunter-gatherers that they could not be genetically related [41]. The comparison of ancient mtDNA
with mtDNA from contemporary Europeans revealed
that there was little genetic similarity between the
hunter-gatherers and modern Europeans, suggesting
that the modern Europeans descended from the
incoming farmers, not from the native populations.
The study suggested that additional waves of migration
and later admixture with hunter-gatherers also
shaped the modern European gene pool. From the
above mentioned, we concluded that there are still
unanswered questions about these populations that
can be solved in future investigations [42].
Another independent research group interested
in Neolithic revolution recently worked with mtDNA
from skeletons of pre Neolithic hunter-gatherers as
well as early Neolithic farmers and compared the
results with modern data. They support the model of
demic diffusion [43]. Researchers identified mtDNA
haplogroups that are typical for early farmers (haplogroup
H) and hunter-gatherers (haplogroup U). The
analyses of mtDNAs typical for hunters and gatherers
showed population expansion between 15,000 and
10,000 years ago and corresponded to an analogous
population increase approximately 9000 years ago,
characterized with mtDNAs typical for early farmers.
The spread of agriculture in Europe pushed the expansion
of farming populations into Europe followed
by the admixture with resident hunter-gatherers. This
was confirmed with results from analyses of mtDNA
belonging to haplo-group H, which are indicative
for population expansions and spread of the farming
culture. The mtDNAs belonging to haplogroup
U showed that the earlier hunter-gatherers adopted
farming practices and admixed with the immigrant
farming populations [44].
Development of Central Europe. In another
study, scientists tried to locate the origins and trace
potential dis-persal routes of populations which had
lived in Central Europe by using comprehensive phylogeographic
and population genetic analyses. They
described the Neolithic mtDNA sequence diversity
and the geographical affinities of the early farmers
using a large database of extant Western Eurasian
and European populations. This was the first detailed
genetic analysis of the earliest Neolithic culture in
Central Europe, which identified Neolithic haplotypes
that left clear traces in the modern populations. The
results confirmed that the demographic events that had
great influence in Europe occurred after the early Neolithic.
These early populations show unique genetic
features and they have an affinity with modern-day
populations from the Near East and Anatolia [45,46].
These data from Central Europe were compared
with the results from the analysis of mtDNA diversity
in hunter-gatherers and first farmers from Northern
Spain. The analyzed samples were dated to the Paleolithic
and Early Neolithic. The results from the analysis
do not support the demic and cultural diffusion
models, but they support a random dispersion model
for Neolithic farmers in the Mediterranean region.
This model points to a differentiation in the various
geographic regions due to the transport of different,
small Neolithic groups from the Near Eastern gene
pool [47].
Research of Past Populations and Their Influence
on Contemporary Ones: Scandinavian Populations.
A research from 2010 concerning Scandinavian
populations was focused on ancient individuals
from different Danish regions dating from the Mesolithic
to the Medieval Age. The results show that
the ancient mtDNA diversity varies in the different
regions. The ancient Danish populations are not so
different from the present-day populations and other
modern Scandinavians, with the exception of haplogroup
I, which is significantly more frequent among
the ancient Danes than among modern Danes and
Scandinavians. The Neolithic and Bronze Age Danish
samples had haplogroups associated with the Mesolithic
populations of Central and Northern Europe (U4
and U5a). These findings show that Southern Scandinavian
populations do not belong to more diverse
haplogroups, which dominated in hunter-gatherer
populations during the Neolithic era [48]. Investigations
made by Helgason et al. [49,50] in the period
from 2003 to 2009, of the early medieval Icelandic
skeletal remains dated from 1000 AD, show that
their sequences are more close to the sequences from
contemporary inhabitants of Scotland, Ireland, and
Scandinavia than to those from the modern Icelandic
population. The scientists thought that this could be
due to the fact that the gene pool of this small Icelandic
population was more affected by genetic drift
(during the last 1100 years) than the mtDNA pools of
the larger European populations [49,50].
Guanches in the Canary Islands. Small-scale
and local anthropogenetic researches can help to
understand the history and to detect the ancestors
of certain populations. Such analyses are those of Rando et al. [51] and Maca-Mayer et al. ]52[, involving
approximately 1000-year-old aboriginal remains
from the Canary Islands. These scientists were
studying the origins of contemporary population of
the Islands, based on historical information from the
prehistoric colonization of the Canary Islands by the
Guanches to the Europeans’ conquest of the Archipelago.
They determined that the Guanches carried
U6b1 mtDNA lineages that are found in the present
day Canary Islands population, but are not found in
Northwest Africans. On other hand, phylogenetical
U6b is not present in the native Canary Islanders.
There is a high genetic and ethnic diversity found in
the Guanches. The comparisons with other populations
related to the Guanches show that despite the
series of population changes, the aboriginal mtDNA
lineages played a big role in the formation of the
Canary Islanders’ gene pool. They concluded that
the Ber-bers are the most probable descendants of
the Guanches, although important human movements
did reshape Northwest Africa after the migratory
wave to the Canary Islands [51,52].
Native Americans. Searching the roots of modern
Native Americans, Kaestle and Smith in 2001
[53] determined the frequencies of five different haplogroups
among the ancient individuals from Western
Nevada dating approximately 350-9200 years before
the present. The Numic speakers (current inhabitants
of the Great Basin, Nevada, North America),
are recent immigrants into the area who replaced the
previous inhabitants. There is a significant genetic
discontinuity between the ancient inhabitants and the
modern Numic speakers that supports the hypothesis
for the Recent Numic Expansion. There is regional
variation over time and it correlates with population
movements hypothesized from linguistics and archaeological
studies. It is also suggested that genetic
drift has not influenced the haplogroup frequency
distributions [53].
Southeast Asia. Investigation of the genetic
relationships and history among two neighboring
populations of early agricultural communities in
northeastern Thailand from the fourth millennium
before the present day and comparing with various
ethnic modern populations of East and Southeast
Asia, shows that the two ancient samples were most
closely related to each other, and closely related
to the part of modern populations living near the
archaeological sites. It was suggested that the genetic
continuum may have persisted since prehistoric
times. Formation of ethnic populations was complicated
culturally and genetically. It was important to
know the contemporary ethnic populations of this
geographic area, direct genetic descendants of their
ethnic and linguistic predecessors, in order to trace
the expansion of the ethnic populations. For the first
time, the analysis of aDNA from these archaeological
sites extends the knowledge about the complex demographic
history and genetic relations of the known
ethnic populations of Southeast Asia [54,55].
Terra Cotta Warriors in China. There is an
interesting analysis from Xu et al. in 2008 [56] performed
with bone remains, which were supposed
to be from workers building the mausoleum for the
First Emperor of China, that were excavated near the
Terra Cotta Warriors and dated 2200 years ago. The
geneticists investigated mitochondrial lineages of
these bone specimens to confirm the hypothesis that
mausoleum-building workers were brought in from
various geographic areas. They saw that the slave
workers of the Qin Dynasty were extremely genetically
diverse. They concluded that the workers were
an admixture and it is difficult to determine genetic
continuity with contemporary Chinese populations.
It is necessary to do more studies in this field [56].
Hungarian Populations. During 2007, the Hungarian
team led by Tomory et al. [57], examined ancient
samples dating back to the 10th-11th centuries
and compared them with contemporary Hungarians
and different European and Asian populations. It was
found that there are genetic differences and no genetic
continuity between ancient and modern Hungarians.
The genetic affinity among ancient Hungarians
and Asian populations is small and the genetic affinity
among ancient Hungarians and Western Eurasian
populations is large. The modern Hungarian populations
are typically European. From the final results,
the conclusion was that Hungarians are genetically
influenced from Slavic, Balkan and Western Eurasian
populations. The Seklers, Hungarian-speaking population
in Transylvania, have more genetic influence
from Eastern and Southern Europeans [57].
Ancient Australians. Adcock et al. in 2001 [58]
made a reconstruction of ancient mtDNA from the
late Pleistocene and early Holocene (from 60,000 to
8000 years ago) eras in Australia. The results indicated
that anatomically modern humans were present in
Australia before the complete fixation of the mtDNA lineages now found in all living people. When comparing
the ancient and modern Australians, scientist
found that the anatomically modern humans were
among those that were replaced and that part of the
replacement occurred in Australia during its colonization.
Other interesting findings were that anatomical
features and the mtDNA of particular individuals
may have had different evolutionary pathways. Some
nuclear gene lineages have different genealogical
and geographical characteristics from mtDNA. The
scientists found that the most divergent mtDNA lineage
from an anatomically modern human was from
an Australian individual. These findings present other
possible models for the demographic and evolutionary
history of our species on the basis of described
deep lineages in Africa and even deeper lineages in
Australia [58].
Yakuts Populations in Siberia. Tracing the origins
of Yakuts, an enigmatic ethnic group living in Siberia,
Crubézy et al. in 2010 [59], extracted mtDNA
from frozen bodies dated between the 15th and 19th
centuries. They used a combination of ethnological
and linguistic criteria as well as population genetic
studies to find their roots. The Yakuts contrast from
other populations in Siberia in different aspects such
as funeral rituals and lifestyles, they are the most
remarkable case of expansion into North Siberia. The
comparison with contemporary populations provided
evidence of microevolution of the Yakut population.
This can be observed in few regions in the world and
it is an interesting scientific finding [59]. The earlier
researches for investigating the origin and evolution
of the Yakut populations performed on frozen bodies
dated 300-600 years ago and older ancient samples
made comparisons with Eurasian modern individuals
and they provided more comprehensive insights of
the Yakut populations history [60].
Native Britons. British history and genetic diversity
are very interesting, and in order to understand
them, Topf et al. [61] performed an analysis of
ancient mtDNA from remains dated between 300 to
1000 years AD, and compared the results with data for
contemporary Britons, Europeans and Middle Eastern
populations. The analysis showed that there is loss
of diversity over the last millennium. The ancient
samples have greater genetic diversity than modern
populations. The observed pattern is probably due
to stochastic processes that lead to the increasing
representation of the ancient haplotypes in modern
populations [61]. Similarly, the gene pool of the Icelandic
population contains evidence of drift distorting
haplotype frequencies as the contribution of ancestral
lines is highly contorted and necessitates further research
to estimate relative importance, because drift
or selection, or both, can be involved [49,61].
Peruvian Populations. Interesting objects of
anthro-pogenetic researches are the Peruvian populations
who lived near Machu Picchu in the Andes in
Peru. They were investigated in 2006 by the team of
Shinoda et al. [62]. The examined populations were
close to populations in the Peruvian and Bolivian
highlands and far from pre Hispanic individuals of
the north coast of Peru. There is a suggestion that late
pre Hispanic individuals and modern Andean highlanders
have strong relations and they were a mixed
group of natives from various coastal and highland
regions relocated by the Inca state. The final results
showed that populations who have lived near to Machu
Picchu are indigenous highlanders who provided
supportive roles for the Machu Picchu region [62].
|
|
|
|
 |
Number 27
VOL. 27 (2), 2024 |
Number 27
VOL. 27 (1), 2024 |
Number 26
Number 26 VOL. 26(2), 2023 All in one |
Number 26
VOL. 26(2), 2023 |
Number 26
VOL. 26, 2023 Supplement |
Number 26
VOL. 26(1), 2023 |
Number 25
VOL. 25(2), 2022 |
Number 25
VOL. 25 (1), 2022 |
Number 24
VOL. 24(2), 2021 |
Number 24
VOL. 24(1), 2021 |
Number 23
VOL. 23(2), 2020 |
Number 22
VOL. 22(2), 2019 |
Number 22
VOL. 22(1), 2019 |
Number 22
VOL. 22, 2019 Supplement |
Number 21
VOL. 21(2), 2018 |
Number 21
VOL. 21 (1), 2018 |
Number 21
VOL. 21, 2018 Supplement |
Number 20
VOL. 20 (2), 2017 |
Number 20
VOL. 20 (1), 2017 |
Number 19
VOL. 19 (2), 2016 |
Number 19
VOL. 19 (1), 2016 |
Number 18
VOL. 18 (2), 2015 |
Number 18
VOL. 18 (1), 2015 |
Number 17
VOL. 17 (2), 2014 |
Number 17
VOL. 17 (1), 2014 |
Number 16
VOL. 16 (2), 2013 |
Number 16
VOL. 16 (1), 2013 |
Number 15
VOL. 15 (2), 2012 |
Number 15
VOL. 15, 2012 Supplement |
Number 15
Vol. 15 (1), 2012 |
Number 14
14 - Vol. 14 (2), 2011 |
Number 14
The 9th Balkan Congress of Medical Genetics |
Number 14
14 - Vol. 14 (1), 2011 |
Number 13
Vol. 13 (2), 2010 |
Number 13
Vol.13 (1), 2010 |
Number 12
Vol.12 (2), 2009 |
Number 12
Vol.12 (1), 2009 |
Number 11
Vol.11 (2),2008 |
Number 11
Vol.11 (1),2008 |
Number 10
Vol.10 (2), 2007 |
Number 10
10 (1),2007 |
Number 9
1&2, 2006 |
Number 9
3&4, 2006 |
Number 8
1&2, 2005 |
Number 8
3&4, 2004 |
Number 7
1&2, 2004 |
Number 6
3&4, 2003 |
Number 6
1&2, 2003 |
Number 5
3&4, 2002 |
Number 5
1&2, 2002 |
Number 4
Vol.3 (4), 2000 |
Number 4
Vol.2 (4), 1999 |
Number 4
Vol.1 (4), 1998 |
Number 4
3&4, 2001 |
Number 4
1&2, 2001 |
Number 3
Vol.3 (3), 2000 |
Number 3
Vol.2 (3), 1999 |
Number 3
Vol.1 (3), 1998 |
Number 2
Vol.3(2), 2000 |
Number 2
Vol.1 (2), 1998 |
Number 2
Vol.2 (2), 1999 |
Number 1
Vol.3 (1), 2000 |
Number 1
Vol.2 (1), 1999 |
Number 1
Vol.1 (1), 1998 |
|
|
