New geochronological, paleoclimatological, and archaeological data from the Narmada Valley hominin locality, central India

The oldest known fossil hominin in southern Asia was recovered from Hathnora in the Narmada Basin, central India in the early 1980's. Its age and taxonomic affinities, however, have remained uncertain. Current estimates place its maximum age at >236 ka, but not likely older than the early middle Pleistocene. The calvaria, however, could be considerably younger. We report recent fieldwork at Hathnora and associated Quaternary type-sections that has provided new geological and archaeological insights. The portion of the exposed ‘Boulder Conglomerate’ within the Surajkund Formation, which forms a relict terrace and has yielded the hominin fossils, contains reworked and stylistically mixed lithic artifacts and temporally mixed fauna. Three mammalian teeth stratigraphically associated with the hominin calvaria were dated by standard electron spin resonance (ESR). Assuming an early uranium uptake (EU) model for the teeth, two samples collected from the reworked surface deposit averaged 49 ± 1 ka (83 ± 2 ka, assuming linear uptake [LU]; 196 ± 7 ka assuming recent uptake [RU]). Another sample recovered from freshly exposed, crossbedded gravels averaged 93 ± 5 ka (EU), 162 ± 8 ka (LU) or 407 ± 21 ka (RU). While linear uptake models usually provide the most accurate ages for this environment and time range, the EU ages represent the minimum possible age for fossils in the deposit. Regardless, the fossils are clearly reworked and temporally mixed. Therefore, the current data constrains the minimum possible age for the calvaria to 49 ± 1 ka, although it could have been reworked and deposited into the Hathnora deposit any time after 160 ka (given the LU uptake ages) or earlier (given the RU ages). At Hathnora, carbonaceous clay, bivalve shells, and a bovid tooth recovered from layers belonging to the overlying Baneta Formation have yielded ¹⁴C ages of 35.66 ± 2.54 cal ky BP, 24.28 ± 0.39 cal ky BP, and 13.15 ± 0.34 ky BP, respectively. Additional surveys yielded numerous lithics and fossils on the surface and within the stratigraphic sequence. At the foot of the Vindhyan Hills 2 km from the river, we recovered a typologically Early Acheulean assemblage comprised of asymmetrical bifaces, large cleavers with minimal working, trihedral picks, and flake tools in fresh condition. These tools may be the oldest Acheulean in the Narmada Valley. Several lithics recovered from the Dhansi Formation may represent the first unequivocal evidence for an early Pleistocene hominin presence in India. In situ invertebrate and vertebrate fossils, pollen, and spores indicate a warm, humid climate during the late middle Pleistocene. High uranium concentrations in the mammalian teeth indicate exposure to saline water, suggesting highly evaporative conditions in the past. Late Pleistocene sediment dated between 24.28 ± 0.39 cal ky BP and 13.15 ± 340 ky BP has yielded pollen and spores indicating cool, dry climatic conditions corresponding to Oxygen Isotope Stage 2 (OIS 2). An early Holocene palynological assemblage from the type locality at Baneta shows evidence for relatively dry conditions and a deciduous forest within the region. The Dhansi Formation provisionally replaces the Pilikarar Formation as the oldest Quaternary formation within the central Narmada Basin. The Baneta Formation, previously dated at 70 ka to 128 ka, correlates with the late Pleistocene and early Holocene. Our results highlight the need for further Quaternary geological and paleoanthropological research within the Narmada Basin, especially because dam construction threatens these deposits.     

The Liang Bua faunal remains: a 95 k.yr. sequence from Flores, East Indonesia

Excavations at Liang Bua, a limestone cave on the island of Flores, East Indonesia, have yielded a well-dated archaeological and faunal sequence spanning the last 95 k.yr., major climatic fluctuations, and two human species - H. floresiensis from 95 to 17 k.yr.¹, and modern humans from 11 k.yr. to the present. The faunal assemblage comprises well-preserved mammal, bird, reptile and mollusc remains, including examples of island gigantism in small mammals and the dwarfing of large taxa. Together with evidence from Early-Middle Pleistocene sites in the Soa Basin, it confirms the long-term isolation, impoverishment, and phylogenetic continuity of the Flores faunal community. The accumulation of Stegodon and Komodo dragon remains at the site in the Pleistocene is attributed to Homo floresiensis, while predatory birds, including an extinct species of owl, were largely responsible for the accumulation of the small vertebrates. The disappearance from the sequence of the two large-bodied, endemic mammals, Stegodon florensis insularis and Homo floresiensis, was associated with a volcanic eruption at 17 ka and precedes the earliest evidence for modern humans, who initiated use of mollusc and shell working, and began to introduce a range of exotic animals to the island. Faunal introductions during the Holocene included the Sulawesi warty pig (Sus celebensis) at about 7 ka, followed by the Eurasian pig (Sus scrofa), Long-tailed macaque, Javanese porcupine, and Masked palm civet at about 4 ka, and cattle, deer, and horse - possibly by the Portuguese within historic times. The Holocene sequence at the site also documents local faunal extinctions - a result of accelerating human population growth, habitat loss, and over-exploitation.     

Astronomical dating of the Xiantai, Donggutuo and Maliang Paleolithic sites in the Nihewan Basin (North China) and implications for early human evolution in East Asia

Magnetostratigraphic studies have established a first-order chronological framework for the Paleolithic sites in the Nihewan Basin (North China), which enabled tracking early human evolution in East Asia. However, to fully understand how well early humans were adapted to climate change, a truly precise dating of the Paleolithic sites is required. Here, we established a high-resolution astronomical timescale for the Xiantai and Donggutuo fluvio-lacustrine successions at the eastern margin of the Nihewan Basin employing low-field magnetic susceptibility (χ) as a climatic indicator, aiming to further refine the ages of the Xiantai, Donggutuo and Maliang Paleolithic sites. Starting from an initial age model constrained by geomagnetic reversals, larger-scale χ cycles were firstly tuned to orbital obliquity using an automatic orbital tuning method. This first-order tuning was followed by simultaneously tuning χ to both obliquity and precession. The finally tuned χ records can be correlated almost cycle-by-cycle with the quartz grain-size record of the Chinese loess sequence and the marine δ¹⁸O record. The astronomically estimated age of the Xiantai Paleolithic site is ca. 1.48 Ma, corresponding to paleosol layer S₂₀ of the Chinese loess sequences or marine oxygen isotope stage (MIS) 49, an interglacial period. The astronomical estimate for the Donggutuo Paleolithic site ranges from ~ 1.06 Ma to 1.12 Ma, corresponding to paleosol/loess layers S₁₁-S₁₂ or MIS 31-33, spanning both interglacial and glacial periods. The astronomically estimated age of the Maliang Paleolithic site is ~ 0.79 Ma, corresponding to loess layer L₈ or MIS 20, a glacial period. This astronomical finding further implies that early humans may have permanently occupied China as far north as 40^oN since at least 1.1 Ma, and before this time the occupation may be intermittent.     

New evidence for a 67,000-year-old human presence at Callao Cave, Luzon, Philippines

Documentation of early human migrations through Island Southeast Asia and Wallacea en route to Australia has always been problematic due to a lack of well-dated human skeletal remains. The best known modern humans are from Niah Cave in Borneo (40-42 ka), and from Tabon Cave on the island of Palawan, southwest Philippines (47 ± 11 ka). The discovery of Homo floresiensis on the island of Flores in eastern Indonesia has also highlighted the possibilities of identifying new hominin species on islands in the region. Here, we report the discovery of a human third metatarsal from Callao Cave in northern Luzon. Direct dating of the specimen using U-series ablation has provided a minimum age estimate of 66.7 ± 1 ka, making it the oldest known human fossil in the Philippines. Its morphological features, as well as size and shape characteristics, indicate that the Callao metatarsal definitely belongs to the genus Homo. Morphometric analysis of the Callao metatarsal indicates that it has a gracile structure, close to that observed in other small-bodied Homo sapiens. Interestingly, the Callao metatarsal also falls within the morphological and size ranges of Homo habilis and H. floresiensis. Identifying whether the metatarsal represents the earliest record of H. sapiens so far recorded anywhere east of Wallace’s Line requires further archaeological research, but its presence on the isolated island of Luzon over 65,000 years ago further demonstrates the abilities of humans to make open ocean crossings in the Late Pleistocene.     

26Al/10Be Burial Dating of Xujiayao-Houjiayao Site in Nihewan Basin,Northern China

The Xujiayao-Houjiayao site in Nihewan Basin is among the most important Paleolithic sites in China for having provided a rich collection of hominin and mammalian fossils and lithic artifacts. Based on biostratigraphical correlation and exploratory results from a variety of dating methods, the site has been widely accepted as early Upper Pleistocene in time. However, more recent paleomagnetic analyses assigned a much older age of ∼500 ka (thousand years). This paper reports the application of ²⁶Al/¹⁰Be burial dating as an independent check. Two quartz samples from a lower cultural horizon give a weighted mean age of 0.24 ± 0.05 Ma (million years, 1σ). The site is thus younger than 340 ka at 95% confidence, which is at variance with the previous paleomagnetic results. On the other hand, our result suggests an age of older than 140 ka for the site’s lower cultural deposits, which is consistent with recent post-infrared infrared stimulated luminescence (pIR-IRSL) dating at 160–220 ka.     

Variabilité et homogénéité des modes de débitage en Chine entre 300 000 et 50 000 ans

The position of lithic industries of Chinese Paleolithic in comparison with those of other regions in the world is always a controversial topic. The typological analysis has made Chinese prehistorians reveal some variability between lithic industries of different regions of China in terms of technical tradition and industrial structure. But this recognition, based upon some morphological characters of stone artifacts, failed to define systematically the modes of production that exist in each site. To answer these problematic some new methods need to be applied. So the lithic materials from 8 Chinese sites dated from 300 ka to 50 ka B.P. were selected and analyzed by means of lithic technological analysis and a new approach for analyzing the structures of cores. This paper presents principally the analytical results of modes of débitage and revealed that the modes of débitage during this period were characterized essentially by system C (“simple débitage”) and E (discoid) and that the concept Levallois was absent during this period in technical systems of lithic production in China. The Chinese lithic industries between 300 ka and 50 ka B.P. saw some homogeneity in the scale of the continent and remarkable difference from those of other regions in the world. So a stable, continuous but independent even isolated evolution in comparison with those of other regions was observed in this period. In addition, despite of apparent homogeneity, some variability exists also between these industries.     

Optically stimulated luminescence dating of cave deposits at the Xiaogushan prehistoric site, northeastern China

The Xiaogushan cave site is one of the most important prehistoric sites in North China. The stone and bone artifacts found in the cave are similar to European contemporaneous artifacts. Cave deposits consist of five layers that have been dated from 46,353 ± 1179 to 4229 ± 135 cal. yr BP, using radiocarbon dating techniques on charcoal and bone samples collected from Layers 2-5. In this paper, optically stimulated luminescence (OSL) techniques were applied to date six samples taken from Layers 1-3. The luminescence properties of the fine-grained and coarse-grained quartz extracts indicate that the materials are suitable for OSL dating using a single-aliquot regeneration-dose (SAR) protocol. The OSL ages obtained are broadly consistent with the stratigraphy and the associated calibrated radiocarbon ages. The dating results show that the cave was first occupied by humans about 70 ka. The human occupation of the cave may be related to climate change. An occupation hiatus is inferred to between ∼17 to ∼10 ka. The stone and bone artifacts found in Layers 2 and 3 may indicate the Middle to Upper Paleolithic transitions in the region.     

Position chronologique des sites de l’Homme moderne en Chine d’après la datation U-Th

The origin and evolution of modern Homo sapiens has been the subject of an intensive debate between exponents of two competing hypotheses, multiregional origins and “recent out of Africa”. This paper presents a synthesis of the chronological studies on seven hominid sites in China based on the U-series dating, five of them of intercalated speleothem calcites and other two of fossil materials. The results show that modern humans were present in China about 100 ka ago, much earlier than previously estimated, and that the so-called “temporal gap” of human presence in China between 40 and 100 ka is most probably nothing but an artifact caused by systematic errors of the dating methods. Further multidisciplinary studies on hominid sites in China may provide important evidence for resolving the hotly debated issues concerning the origin of modern humans.     

Palaeoenvironmental studies in NW Iberia (Cantabrian range): Vegetation history and synthetic approach of the last deglaciation phases in the western Mediterranean

At the north-western limit of the Mediterranean basin, in the Mediterranean-Oceanic transition zone, the multidisciplinary study of two palaeolakes: Villaseca and La Mata (Province of Leon), provides data on the last deglaciation. The juxtaglacial lakes were formed during a deglaciation phase of the last cold stage (Weichselian/Würm) estimated, from the regional and Pyrenean chronological data, between circa 48 ka and 32 ka (ka: 1000 years ago). Their drainage occurred circa 17 ka and 15 ka respectively, during the definitive glacial retreat phases. These results provide evidence for a glacial maximum extension preceding the global Last Glacial Maximum (LGM) and an early deglaciation. Comparisons with the Pyrenean data show that in the most important glacial basins of the Pyrenean-Cantabrian range, glaciers were still present at middle elevation circa 18-20 ka. The final deglaciation did not happen until around 16 ka.During the period 48-13.3 ka, along the Pyrenean-Cantabrian range, three stages can be distinguished in the deglacial processes. Between circa 48 and 32 ka, the deglacial retreats were contemporaneous with D-O events 12 to 6. Between circa 32 ka and 18 ka, major sedimentary modifications were rare. Two of them are synchronous with D-O 4 and 3. A glacial readvance also occurred during this period along the Pyrenean-Cantabrian range. Between 18 and 17 ka, major sedimentary changes and an increase in the δ¹⁸O records of the NGRIP ice-core record are synchronous. Between 15 and 13.3 ka, glacial retreats and sedimentary changes are synchronous with the climatic improvement observed from the end of the Oldest Dryas to the end of the Bölling-Alleröd period.These correspondencies suggest relationships between the deglacial processes in the western Mediterranean and the palaeoenvironmental events in the North Atlantic region during the last Glacial-Interglacial transition. During the Full Glacial period, the pollen data indicates dominant steppic environments. The Oldest Dryas was not a constant cooling phase and from around 15.5 ka, the pollen data indicates a slight warming. During the Holocene, until circa 2040 cal BP, mixed oak forests developed and pine was a prominent component of the Cantabrian landscape. Then the forests were destroyed due to mining, agriculture and fire, and this favoured the extension of heathland.     

Holocene space-time succession of the Middle Atrato wetlands, Chocó biogeographic region, Colombia

Successional sequences from forested wetlands in the Middle Atrato River Basin were reconstructed using characterisation of present vegetation communities and palynological analysis. A 4.8 km transect, drawn across a river meander, and two 6 and 8 m deep sediment cores (San Martín and Villanueva) were collected in the floodplain within two different vegetation assemblages. Based on the floristic and environmental characteristics of the local vegetation communities, ecological changes spanning the last 4 ka (cal years BP) were analysed in San Martín and Villanueva cores. Present vegetation is dominated by four communities determined by flood tolerance and drainage conditions. We found Euterpe oleraceae, Mauritiella macroclada-Campnosperma panamensis and Oenocarpus bataua forests, and mixed forest and open vegetation in a gradient from poor to improved drainage conditions. Vegetation changes in the palynological record suggest that sedimentation and erosion processes on flood basins are due to changes in drainage conditions and to variable flooding levels.A wet period in the 4 to 2.7 ka interval is postulated, which might be related to sea level rise or local subsidence. Lower flooding levels and improved drainage conditions dominated the 2.7 to ∼ 1.6 ka interval, whereas a flooding event (and a hiatus) occurred between 1.5 and 0.5 ka. This flooding event might be synchronous with analogous events as recorded in the Colombian Amazonia between 1.6 and 1.45 ka. Forest disturbance, probably of anthropogenic origin, is recorded in both sites since 0.5 ka.     

Reconstructing the geomorphic history of Liang Bua, Flores, Indonesia: a stratigraphic interpretation of the occupational environment

Liang Bua, in Flores, Indonesia, was formed as a subterranean chamber over 600 ka. From this time to the present, a series of geomorphic events influenced the structure of the cave and cave deposits, creating a complex stratigraphy. Within these deposits, nine main sedimentary units have been identified. The stratigraphic relationships between these units provide the evidence needed to reconstruct the geomorphic history of the cave. This history was dominated by water action, including slope wash processes, channel formation, pooling of water, and flowstone precipitation, which created waterfalls, cut-and-fill stratigraphy, large pools of water, and extensive flowstone cappings. The reconstructed sequence of events over the last 190 k.yr. has been summarized by a series of time slices that demonstrate the nature of the occupational environment in Liang Bua. The earliest artifacts at the site, dated to ∼190 ka, testify to hominin presence in the area, but the reconstructions suggest that occupation of the cave itself may not have been possible until after ∼100 ka. At ∼95 ka, channel erosion of a basal unit, which displays evidence of deposition in a pond environment, created a greater relief on the cave floor, and formed remanent areas of higher ground that later became a focus for hominin occupation from 74-61 ka by the west wall and in the center of the cave, and from ∼18-17 ka by the east wall. These zones have been identified according to the sloping nature of the stratigraphy and the distribution of artifacts, and their locations have implications for the archaeological interpretation of the site.     

A 2700-year high resolution pollen record of climate change from varved Sugan Lake in the Qaidam Basin, northeastern Tibetan Plateau

A 2700-year high resolution pollen record from annually-varved Sugan Lake in the Qaidam Basin at 2793 m a.s.l was obtained to examine vegetation and climatic change on the NE Tibetan Plateau. Pollen data shows that Sugan Basin was constantly covered by open desert-steppe vegetation dominated by Chenopodiaceae, Artemisia, Poaceae and Ephedra. However, large variations in Artemisia/Chenopodiaceae (A/C) ratios suggest regional moisture fluctuations over the last 2700 years, including a dry and relatively stable climate prior to 300 AD, relatively wet climate from 300 to 1200 AD with variability during 1100-1200 AD, and unstable climate since 1200 AD with relatively moister climate during 1250-1400 AD and 1700-1800 AD. However, other proxies (varve thickness, Chironomid taxa, isotopes of oxygen in precipitated carbonate) show fresher water when regional moisture was lower inferred from A/C ratio. This inconsistency suggests the possible difference of in-lake lithology/environment and regional moisture change. Fresh water into the lake from ice melting on the surrounding mountains might have contributed to the in-lake lithology and environment variation. The effective moisture changes in the Basin are in opposite phases to snow accumulation records from Dunde ice core (5325 m a.s.l) and to the monsoon intensity inferred from Dongge Cave, suggesting that the regional topography might have played an important role in mediating moisture changes at regional scale. Pollen data from Sugan Lake shows the shift of moisture at 1200 AD, from stable to variable conditions. This event is well correlated with other paleoclimate proxies in China and other parts of the world; however, the mechanisms behind these patterns require further investigation.     

The evolving landscape and climate of western Flores: an environmental context for the archaeological site of Liang Bua

The rapidly changing landscape of the eastern Indonesian archipelago has evolved at a pace dictated by its tropical climate and its geological and tectonic history. This has produced accelerated karstification, flights of alluvial terraces, and complex, multi-level cave systems. These cave systems sometimes contain a wealth of archaeological evidence, such as the almost complete skeleton of Homo floresiensis found at the site of Liang Bua in western Flores, but this information can only be understood in the context of the geomorphic history of the cave, and the more general geological, tectonic, and environmental histories of the river valley and region. Thus, a reconstruction of the landscape history of the Wae Racang valley using speleothems, geological structure, tectonic uplift, karst, cave, and terrace development, provides the necessary evidence to determine the formation, age, evolution, and influences on the site. This evidence suggests that Liang Bua was formed as two subterranean chambers ∼600 ka, but could not be occupied until ∼190 ka when the Wae Racang wandered to the southern side of the valley, exposing the chamber and depositing alluvial deposits containing artifacts. During the next ∼190 k.yr., the chambers coalesced and evolved into a multi-level and interconnected cave that was subjected to channel erosion and pooling events by the development of sinkholes. The domed morphology of the front chamber accumulated deep sediments containing well stratified archaeological and faunal remains, but ponded water in the chamber further prevented hominin use of the cave until ∼100 ka. These chambers were periodically influenced by river inundation and volcanic activity, whereas the area outside the cave was greatly influenced by glacial phases, which changed humid forest environments into grassland environments. This combined evidence has important implications for the archaeological interpretation of the site.     

Remediation of copper-contaminated soil by Kocuria flava CR1, based on microbially induced calcite precipitation

An indigenous calcifying bacterial strain CR1, identified as Kocuria flava, was isolated from soil of a mining area, Urumqi, China. An extensive copper bioremediation capacity of this isolate was studied based on microbially induced calcite precipitation (MICP). K. flava CR1 removed 97% of copper when initial Cu concentration was 1000 mg L⁻¹. The isolate produced significant amount of urease (472 U mL⁻¹), an enzyme that leads to calcite precipitation. The isolate removed 95% of copper from contaminated soil. The MICP process in bioremediation was further confirmed by FTIR and XRD analyses. FTIR analysis showed two different forms of calcium carbonate, i.e., calcite and aragonite, and the results were well supported by XRD. For the first time, the ability of K. flava has been documented in the bioremediation of polluted soil. This study showed that MICP-based bioremediation by K. flava is a viable, environmental friendly technology for cleaning-up the copper-contaminated site.     

Mass spectrometric U-series dating of Huanglong Cave in Hubei Province,central China: Evidence for early presence of modern humans in eastern Asia

Most researchers believe that anatomically modern humans (AMH) first appeared in Africa 160-190 ka ago, and would not have reached eastern Asia until ∼50 ka ago. However, the credibility of these scenarios might have been compromised by a largely inaccurate and compressed chronological framework previously established for hominin fossils found in China. Recently there has been a growing body of evidence indicating the possible presence of AMH in eastern Asia ca. 100 ka ago or even earlier. Here we report high-precision mass spectrometric U-series dating of intercalated flowstone samples from Huanglong Cave, a recently discovered Late Pleistocene hominin site in northern Hubei Province, central China. Systematic excavations there have led to the in situ discovery of seven hominin teeth and dozens of stone and bone artifacts. The U-series dates on localized thin flowstone formations bracket the hominin specimens between 81 and 101 ka, currently the most narrow time span for all AMH beyond 45 ka in China, if the assignment of the hominin teeth to modern Homo sapiens holds. Alternatively this study provides further evidence for the early presence of an AMH morphology in China, through either independent evolution of local archaic populations or their assimilation with incoming AMH. Along with recent dating results for hominin samples from Homo erectus to AMH, a new extended and continuous timeline for Chinese hominin fossils is taking shape, which warrants a reconstruction of human evolution, especially the origins of modern humans in eastern Asia.     

A new elasmothere (Perissodactyla, Rhinocerotidae) from the late Miocene of the Linxia Basin in Gansu, China

Ningxiatherium euryrhinus sp. nov. is a relatively large elasmothere rhino from the Linxia Basin in northwestern China found in the early Bahean, which corresponds to the early Vallesian (MN 9), ca. 11.1 Ma. It is much larger than the extant Ceratotherium simum and has a single horn. Ningxiatherium is similar to the late Miocene Parelasmotherium from Gansu and Shanxi, but differs in having partially ossified nasal septum, terminal nasal horn boss, shallower nasal notch above the P3/P4 boundary, and much shorter premolars. N. euryrhinus sp. nov. differs from N. longirhinus from the late Miocene of Zhongning, Ningxia in northwestern China by its larger size, relatively wider nasals, shallower nasal notch, and sub-quadrangular occlusal surface of M3. In the Linxia Basin, the other early late Miocene (Bahean or Vallesian) elasmotheres are known from complete skulls or isolated teeth. They include two genera and three species: Parelasmotherium linxiaense, P. simplum, and Iranotherium morgani. They lack a nasal septum, but Ningxiatherium has one. The age of N. euryrhinus is early late Miocene based on direct association with biochronologic indicator taxa, such as Dinocrocuta gigantea, Hipparion dongxiangense, and Chilotherium primigenius. N. euryrhinus is more primitive than N. longirhinus sp. nov. in having a sub-quadrangular M3, a shallower nasal notch, and the presence of DP1 in adults.     

Preface: research at Liang Bua, Flores, Indonesia

Excavations at Liang Bua, Flores, Indonesia, have yielded evidence for an endemic human species, Homo floresiensis, a population that occupied the cave between ∼95-17 ka. This discovery has major implications for early hominin evolution and dispersal in Africa and Asia, attracting worldwide interest. This preface describes the rationale for the excavations in historical, geographical, and wider research contexts, as well as the methods used. It also introduces the other papers on aspects of Liang Bua research that feature in this edition of the Journal of Human Evolution.     

Spectral resolution of a second binding site for Nile Red on cytochrome P4503A4

Nile Red is sequentially metabolized by cytochrome P4503A4 to the N-monoethyl and N-desethyl products, which typifies the metabolism of many amine containing drugs. Sequential metabolism of a single substrate results in complex kinetics that confound predictive models of drug clearance. As a fluorescent model for drugs which undergo sequential metabolism, Nile Red provides the opportunity to monitor drug-CYP interactions wherein the fluorescent properties of Nile Red could, in principle, be exploited to determine individual rate and equilibrium constants for the individual reactions. Previously, it was shown that Nile Red binds at the active site and fluoresces (K_D ≈ 50 nM) with maximum emission at ∼620 nm, but it was unclear whether a red-shifted emission, at ∼660 nm, consisted of only free Nile Red or Nile Red bound at a second site on the protein. Here, equilibrium binding studies, including ‘reverse titrations’ spanning low ratios of CYP3A4/Nile Red, indicate two binding sites for Nile Red with a contribution to the ‘red emission’ greater than can be accounted for by free Nile Red. Singular value decomposition affords basis spectra for both spectral components and fits well to the experimentally determined concentration dependence of Nile Red emission. In addition, the red spectral component, with an apparent K_D = 2.2 μM, is selectively eliminated by titration with the known allosteric effectors of CYP3A4, α-napthoflavone and testosterone. Furthermore, the double mutant L2311F/D214E, previously demonstrated to perturb a peripheral allosteric site, lacks the red-emitting Nile Red binding site, but retains the blue-emitting site. Together these data indicate that a second Nile Red site competes with other effectors of CYP3A4 at a site that results in Nile Red emission at 660 nm.     

Stratigraphie et chronologie du site de Gaolingpo dans le bassin de Bose,sud de la Chine

The Bose Paleolithic industry in the Bose Basin, Guangxi, South China is well known in the international scientific community for its bearing handaxes dated back to 803 ka. Seven terraces were developed in the basin and terrace 4 is the most important because handaxes and tektites were recovered from this terrace. Since the first site was discovered in 1973, additional sites of the Bose Paleolithic industry have been investigated and excavated, most of which are located on terrace 4. The previous studies concluded that there is only one phase within this industry and its age is 803 ka. However, before 2013, no attempt had been made to dig down to the basal gravels from the top of terrace 4 in archaeological excavations. Therefore, the stratigraphy and the layers from which the stone artifacts derive of terrace 4 are not clear. In 2013–2014, we conducted an excavation of 200 m² at the Gaolingpo site, which is one of the most important Paleolithic sites in the Bose Basin. We excavated to the basal gravel deposits from the top of this terrace, and a section of more than 7 meters thick deposit was exposed and clearly shows the stratigraphic layers of Pleistocene and Holocene deposits. Furthermore, features and about 800 stone artifacts including choppers, picks, scrapers, etc. were excavated from several stratigraphic layers, which are different in age and other aspects. Based on stratigraphy and the stone artifacts, three stages of Paleolithic cultural remains from the Gaolingpo site can be established. The first stage is in the age of at leat 803 ka. The second stage can be dated to 15 ka and the third stage to about 10 ka.     

Lactose Binding to Galectin-1 Modulates Structural Dynamics, Increases Conformational Entropy, and Occurs with Apparent Negative Cooperativity

Galectins are a family of lectins with a conserved carbohydrate recognition domain that interacts with β-galactosides. By binding cell surface glycoconjugates, galectin-1 (gal-1) is involved in cell adhesion and migration processes and is an important regulator of tumor angiogenesis. Here, we used heteronuclear NMR spectroscopy and molecular modeling to investigate lactose binding to gal-1 and to derive solution NMR structures of gal-1 in the lactose-bound and unbound states. Structure analysis shows that the β-strands and loops around the lactose binding site, which are more open and dynamic in the unbound state, fold in around the bound lactose molecule, dampening internal motions at that site and increasing motions elsewhere throughout the protein to contribute entropically to the binding free energy. CD data support the view of an overall more open structure in the lactose-bound state. Analysis of heteronuclear single quantum coherence titration binding data indicates that lactose binds the two carbohydrate recognition domains of the gal-1 dimer with negative cooperativity, in that the first lactose molecule binds more strongly (K₁ = 21 ± 6 × 10³ M^− 1) than the second (K₂ = 4 ± 2 × 10³ M^− 1). Isothermal calorimetry data fit using a sequential binding model present a similar picture, yielding K₁ = 20 ± 10 × 10³ M^− 1 and K₂ = 1.67 ± 0.07 × 10³ M^− 1. Molecular dynamics simulations provide insight into structural dynamics of the half-loaded lactose state and, together with NMR data, suggest that lactose binding at one site transmits a signal through the β-sandwich and loops to the second binding site. Overall, our results provide new insight into gal-1 structure-function relationships and to protein-carbohydrate interactions in general.