Establishing a minimum PMI for bone sun bleaching in a UK environment with a controlled desert-simulated comparison.

Microenvironments play a significant part in understanding the post-mortem interval in forensic taphonomy. Recently, the value of weathering factors in relation to obtaining a PMI has been investigated further. In this study, observations were made to calculate the length of time it takes for three different bone elements (femur, rib, and scapula) to bleach in a UK summer and winter. This research also investigated whether there were any physicochemical modifications to the bone caused by bleaching. Porcine femora, scapulae, and ribs were placed into open and shaded areas of an outdoor research facility located in Oxfordshire, UK, during summer (July-Sep) and winter months (Dec-Mar). The specimens were monitored at 3-week intervals using photography, and an observational scoring method was developed to quantify the extent of bleaching. As temperatures are typically much lower in the UK compared with warmer climates, a controlled indoor-simulated desert experiment was also undertaken to be used as a control. This allowed sun bleaching and changes to the bone chemistry to be monitored in a controlled, high-UV environment for comparison with the UK outdoor experiments. Fourier transform infrared spectroscopy (FTIR) was employed to analyze physicochemical modifications to both the mineral and organic components of the bone. The FTIR was used to calculate crystallinity index (CI), mineral to organic ratio, and the relative amount of carbonate concentrations. Weather data was collected and a positive correlation was found between both ultraviolet (UV) levels and accumulated degree days (ADD) when compared with observational bleaching scores. Bleaching (whitening) of the bone samples occurred in both seasons but at different rates, with the bleaching process occurring at a slower rate in winter. During summer, the initial bleaching process was evident at 6 weeks, and by 9 weeks, the bones were an off-white colour. During the winter period, whitening of the bone started at 9 weeks; however, only the scapula and rib samples displayed a similar off-white colour. This colouration was observed at 13 weeks rather than at 9 weeks. The desert simulation samples started bleaching in a similar pattern to the outdoor samples after 1 week but the bones did not fully bleach. The bone chemistry, based on physicochemical properties obtained from the FTIR, showed a significant statistical difference between the simulated desert and winter season when compared against a control sample. For the winter samples, the mineral to organic ratio was significantly higher than that in the control, suggesting a reduction in the proportion of organic. For the samples in the simulated desert environment, the crystallinity index was significantly higher than that in the control samples, suggesting an increase in crystallinity. The results of this experiment support the fact that it is possible to achieve bleaching in a UK environment and that the minimal time frame for this to occur differs in seasons.

Diet and adult age-at-death among mobile foragers: A synthesis of bioarcheological methods.

OBJECTIVES: The research explores whether the combined study of cortical bone histology, bone morphology, and dietary stable isotopes can expand insights into past human health and adaptations, particularly dietary sufficiency and life span. MATERIALS AND METHODS: Midthoracic rib cortices from 54 South African Late Holocene adult skeletons (28 M, 24 F, two sex undetermined) are assessed by transmitted-light microscopy for cross-sectional area measurements, osteon area (On.Ar), osteon population density, and presence/absence of secondary osteon variants. Values for δ13 Cbone collagen , δ15 Nbone collagen , 14 C dates, Southwestern and Southern Cape geographic regions, body size measures, estimated ages-at-death from both morphological and histological methods are integrated into analyses, which include Spearman correlations, χ2 tests and Kruskal-Wallis ANOVAs. RESULTS: There is reduced On.Ar variability with higher δ15 N (r = -.41, p = .005); rib %cortical area and δ15 N are negatively correlated in the Southern Cape group (r = -.60, p = .03). Osteon variants are more common in older adults; histological ages at death are significantly older than those determined from gross morphology. DISCUSSION: We found bone tissue relationships with measures of diet composition, but indicators of dietary adequacy remain elusive. Relationships of tissue quality and isotopes suggest that some Southern Cape adults lived long lives. Osteon variants are associated with age-at-death; some association with diet remains possible. Gross morphological methods appear to underestimate adult ages-at-death, at least among small-bodied adults.

Stable isotope study on ancient populations of central sudan: Insights on their diet and environment.

OBJECTIVES: A contribution to the knowledge of the economy and the environmental surroundings of the populations living along the Nile valley in three different periods. MATERIALS AND METHODS: This study reports stable isotope analyses on apatite bone samples of 139 humans, 48 mammals, and 43 fish from the Al Khiday archaeological sites in Sudan. The bones belong to four archaeological periods: pre-Mesolithic, Mesolithic, Neolithic, and Meroitic. Data were processed statistically. RESULTS: A significant difference exists between the pre-Mesolithic and Mesolithic mean δ(18) Ow value and the mean of the modern Nile. The mean δ(18) Ow values for the Neolithic humans and bovids are very similar (+1.5‰ ±4‰, and -2‰, respectively) and more positive than the mean values of Mesolithic mammals and Pre-Mesolithic humans. The water ingested by Meroitic humans (+7‰ ± 2.5‰) is enriched in (18) O in respect to the water ingested by the Neolithic population. There is a separation in the δ(13) Cdiet values between the pre-Mesolithic humans (-14‰ ± 1‰) and Mesolithic mammals (-12‰ ± 2‰) group and the Neolithic humans (-18‰ ± 1‰), Meroitic humans (-19‰ ±1‰), Neolithic mammals (-21‰), and the modern (mean δ(13) Cdiet = -19‰ ±2‰) mammal group. DISCUSSION: The climate became warmer and more arid from the pre-Mesolithic/Mesolithic to the Meroitic period. The environmental conditions influenced the strategies of subsistence and, in particular, the changes occurring from the pre-Mesolithic to the Neolithic can be considered contemporaneous to the transition from hunting-gathering-fishing to cultivation-herding. Am J Phys Anthropol 160:498-518, 2016. © 2016 Wiley Periodicals, Inc.

Infant feeding practices and childhood diet at Apollonia Pontica: Isotopic and dental evidence.

OBJECTIVE: This study analyzes deciduous dental pathology and stable isotopes to investigate the relationship between diet, feeding practices, and oral health in a subadult skeletal sample from the Greek colonial site of Apollonia Pontica, Bulgaria (mid-5th to mid-3rd Centuries BC). METHODS: Stable carbon and nitrogen isotope analysis was conducted on 74 bone collagen samples, and the deciduous dentitions of 85 individuals aged 8.5 months to 11 years were examined for evidence of caries, calculus, antemortem tooth loss, abscesses, and occlusal tooth wear. RESULTS: δ(13) C and δ(15) N values of the collagen samples indicate that weaning began between the ages of 6 months and 1 year, and was complete for most individuals by the age of 3. The isotopic data are consistent with a mixed diet of primarily terrestrial C3 resources. The dental pathology data indicate that complementary foods provided to young children had an impact on their oral health early on. Four outliers exhibited elevated δ(15) N values compared with the adult female range and lower levels of tooth wear than other members of their age groups. Possible explanations include prolonged breastfeeding, the consumption of diets elevated in (15) N, and physiological/nutritional stress. CONCLUSIONS: The deciduous dental data correlate well with the isotopic data and are consistent with later textual sources regarding the timing and duration of weaning, and the composition of complementary foods. The results of this research demonstrate the value of combining isotopic and dental evidence to investigate the dietary practices of infants and young children and the impact of these practices on oral health.

Oxygen isotope analysis of human bone phosphate evidences weaning age in archaeological populations.

Here we report bone phosphate oxygen (δ(18)Op) values from perinates/neonates and infants (<3.5 years; n = 32); children (4-12 years; n = 12); unsexed juveniles (16-18 years; n = 2); and adult bones (n = 17) from Wharram Percy, North Yorkshire, England, in order to explore the potential of this method to investigate patterns of past breastfeeding and weaning. In prior studies, δ(15)N and δ(13)C analyses of bone collagen have been utilized to explore weaning age in this large and well-studied assemblage, rendering this material highly appropriate for the testing and development of this alternative method targeting the inorganic phase of bone. Data produced reveal (18)O-enrichment in the youngest perinatal/neonatal and infant samples, and an association between age and bone δ(18)Op (and previously-published δ(15)N values), with high values in both these isotope systems likely due to breastfeeding. After the age of 2-3 years, δ(18)Op values are lower, and all children between the ages of 4 and 12, along with the vast majority of sub-adults and adults sampled (aged 16 to >50 years), have δ(18)Op values consistent with the consumption of local modern drinking water. The implications of this study for the reconstruction of weaning practices in archaeological populations are discussed, including variations observed with bone δ(15)Ncoll and δ(18)Op co-analysis and the influence of culturally-modified drinking water and seasonality. The use of this method to explore human mobility and palaeoclimatic conditions are also discussed with reference to the data presented.

Weaning age in an expanding population: stable carbon and nitrogen isotope analysis of infant feeding practices in the Okhotsk culture (5th-13th centuries AD) in Northern Japan.

OBJECTIVE: The Okhotsk people were sedentary hunter-gatherer-fishers who lived and prospered in Sakhalin, Hokkaido, and the Kurile Islands during the 5th to 13th centuries AD. They expanded rapidly along the northeastern coast of Hokkaido. We reconstructed infant feeding practices of the Moyoro population of the Okhotsk culture in eastern Hokkaido, Japan. METHODS: Stable isotope ratios in 58 subadult human skeletons were measured. RESULTS: The results suggest that complementary foods with a relatively low carbon isotope ratio were consumed during and after weaning, as observed in ethnographic descriptions of northern human populations such as the Ainu and isotopically suggested in ancient northern hunter-gatherer-fisher populations. Nitrogen isotope ratios of subadults showed that the age at the end of weaning in the Moyoro population was 1.8 (1.4-2.2 in 95% credible interval) years, which is earlier than that in other northern hunter-gatherer-fisher populations. CONCLUSIONS: Because weaning age is one of the most important determinants of fertility, a shorter breastfeeding period suggests increased fertility. Furthermore, better nutrition would further promote the population increase, and thus populations of the Okhotsk culture could expand into new regions. These findings are consistent with recent emerging evidence of great contributions of the Okhotsk to the formation of later Ainu populations and culture.

Identification of the geographical place of origin of an unidentified individual by multi-isotope analysis.

A multi-isotope investigation (Sr and Pb isotopes and δ18O, δ13C and δ15N) was applied to bone and teeth from an unidentified male found drowned in the"IJ" Ruyterkade in Amsterdam, The Netherlands in March of 1999. The individual remained unidentified until mid 2013, after the isotope study was completed. Coupled δ13C and δ15N values in bone collagen recovered from rib and femur are consistent with an omnivore living in a region where C3-type diet dominates (i.e. Europe). Integrated Sr and Pb isotopes and δ18O values in canine and third molar teeth and femur and rib bone data exclude extended residence in north-west Europe and particularly The Netherlands. Characteristic Pb isotope ratios coupled with inferred δ18O values of drinking water argue for a most probable place of origin for the unidentified individual in west and south Poland, south-east Slovakia and the region of Ukraine-Romania-Bulgaria, specifically the region associated with the Carpathian Mountains. Independent of the isotope study, the Cold Case Team made a positive identification with an individual from south-west Poland, validating the results of the multiple-isotopic approach.

Biochemical properties of tissue-engineered cartilage.

OBJECTIVE: Microtia is treated with rib cartilage sculpting and staged procedures; though aesthetically pleasing, these constructs lack native ear flexibility. Tissue-engineered (TE) elastic cartilage may bridge this gap; however, TE cartilage implants lead to hypertrophic changes with calcification and loss of flexibility. Retaining flexibility in TE cartilage must focus on increased elastin, maintained collagen II, decreased collagen X, with prevention of calcification. This study compares biochemical properties of human cartilage to TE cartilage from umbilical cord mesenchymal stem cells (UCMSCs). Our goal is to establish a baseline for clinically useful TE cartilage. METHODS: Discarded cartilage from conchal bowl, microtic ears, preauricular tags, rib, and TE cartilage were evaluated for collagen I, II, X, calcium, glycosaminoglycans, elastin, and fibrillin I and III. Human UCMSCs were chondroinduced on 2D surfaces and 3D D,L-lactide-co-glycolic acid (PLGA) fibers. RESULTS: Cartilage samples demonstrated similar staining for collagens I, II, and X, elastin, and fibrillin I and III, but differed from rib. TE pellets and PLGA-supported cartilage were similar to auricular samples in elastin and fibrillin I staining. TE samples were exclusively stained for fibrillin III. Only microtic samples demonstrated calcium staining. CONCLUSIONS: TE cartilage expressed similar levels of elastin, fibrillin I, and collagens I and X when compared to native cartilage. Microtic cartilage demonstrated elevated calcium, suggesting this abnormal tissue may not be a viable cell source for TE cartilage. TE cartilage appears to recapitulate the embryonic development of fibrillin III, which is not expressed in adult tissue, possibly providing a strategy to control TE elastic cartilage phenotype.

Stable isotope canopy effects for sympatric monkeys at Tai Forest, Cote d'Ivoire.

This study tests the hypothesis that vertical habitat preferences of different monkey species inhabiting closed canopy rainforest are reflected in oxygen isotopes. We sampled bone from seven sympatric cercopithecid species in the Taï forest, Côte d'Ivoire, where long-term study has established taxon-specific patterns of habitat use and diet. Modern rib samples (n = 34) were examined for oxygen (δ(18)Oap) and carbon (δ(13)Cap) from bone apatite ('bioapatite'), and carbon (δ(13)Cco) and nitrogen (δ(15)Nco) from bone collagen. Results are consistent for C3 feeders in a closed canopy habitat. Low irradiance and evapotranspiration, coupled with high relative humidity and recycled CO2 in forest understory, contribute to observed isotopic variability. Both δ(13)Cco and δ(13)Cap results reflect diet; however, δ(13)C values are not correlated with species preference for canopy height. By contrast, δ(18)Oap results are correlated with mean observed height and show significant vertical partitioning between taxa feeding at ground, lower and upper canopy levels. This oxygen isotope canopy effect has important palaeobiological implications for reconstructing vertical partitioning among sympatric primates and other species in tropical forests.

Stable isotope ratio analysis of breastfeeding and weaning practices of children from medieval Fishergate House York, UK.

Rib collagen of 51 juveniles and 11 adult females from the late medieval Fishergate House cemetery site (York, UK) were analyzed using nitrogen and carbon stable isotope ratio analysis to determine the weaning age for this population and to reconstruct diet. The juveniles' ages ranged from fetal to 5-6 years, while the females were of reproductive age. Previous researchers suggested that the children from Fishergate House might have been weaned later than the medieval British norm of 2 years, based on a mortality peak at 4-6 years of age. The results show weaning was complete by 2 years of age, agreeing with previous British weaning studies. The adult female δ(15) N values have a mean of 11.4‰ ± 1.1‰ and the δ(13) C values have a mean of -19.4‰ ± 0.4‰. These findings are consistent with previous isotopic studies of female diet in York during this period, though slightly lower. The weaned juvenile nitrogen values were found to be higher than the adult females (12.4‰ ± 1.0‰ for δ(15) N and -19.7‰ ± 0.5‰ for δ(13) C), which might indicate a dependence on higher trophic level proteins such as marine fish or pork. Marine fish is considered a high status food and children are considered low-status individuals at this time, making this a particularly interesting finding. Weaning does not appear to coincide with peak mortality, suggesting environment factors may be playing a larger role in child mortality at Fishergate House.

Data for the Reference Man: skeleton content of chemical elements.

This study was undertaken to provide reference values of chemical element mass fractions in intact bone of Reference (European Caucasian) Man/Woman. The rib bone samples investigated were obtained from autopsies of 84 apparently healthy 15-58-year-old citizens (38 females and 46 males) of a non-industrial region in the Central European part of Russia who had suffered sudden death. The mass fractions (mg/kg given on a wet mass basis) of 69 elements in these bone samples were measured by using neutron activation analysis with high-resolution spectrometry of short-lived and long-lived radionuclides, particle-induced gamma-ray emission, inductively coupled plasma atomic emission spectrometry, and inductively coupled plasma mass spectrometry including necessary quality control measures. Using published and measured data, mass fraction values of the 79 elements for the rib bone have been derived. Based on accepted rib to skeleton mass fractions and reference values of skeleton mass for Reference Man, the elemental burdens in the skeleton were estimated. These results may provide a representative bases for establishing related reference values for the Russian Reference Man/Woman and for revising and adding current reference values for the International Commission on Radiological Protection. The data presented will also be very valuable for many other applications in radiation protection, radiotherapy radiation dosimetry, and other scientific fields.

Mechanical mechanism of rib spalling and sensitivity analysis of gangue parameters to rib spalling in gangue-bearing coal seams.

Rib spalling is one of the main factors restricting the safe and efficient production of the fully mechanized mining face in gangue-bearing coal seams, and the gangue has significant influence on the occurrence of rib spalling. In this study, the instability process and mechanical mechanism of rib spalling in gangue-bearing coal seams were studied, and the sensitivity of gangue parameters to rib spalling was analyzed. The simulation test of rib spalling under different gangue parameters was carried out by orthogonal tests. The width and depth of rib spalling were taken as evaluation indexes, and the influence of gangue parameters on the rib spalling was analyzed by variance analysis and significance tests. The results show that the failure process of rib spalling is characterized by the fracturing failure of the lower coal body, shear failure of the gangue layer, and the falling off of the upper coal body caused by the gravity; the gangue parameters (thickness, density, joint inclination, and internal friction angle) have an important influence on the sliding instability of the coal wall. In the sensitivity analysis, the influence of gangue parameters on the width of rib spalling is ordered as gangue density > joint inclination > gangue thickness > internal friction angle; the influence of gangue parameters on the depth of rib spalling is ordered as gangue density > joint inclination > internal friction angle > gangue thickness. Besides, the greater the gangue density, the less damage caused by stress concentration, and the lower the risk of rib spalling.

Biochemical Compositional Analysis of Cadaveric Rib Cartilage.

Background: Cadaveric rib is used as a cartilage source for reconstructive rhinoplasty in patients who lack sufficient native septal cartilage; however, these grafts are known to warp. Objective: To measure and compare the biochemical properties of cadaveric rib as related to age, gender, and cortical versus core location. Methods: Seven cadaveric rib cartilage specimens were obtained and sectioned into cortical and core segments. Biochemical assays were used to determine total collagen and sulfated glycosaminoglycan (sGAG) content. Results: Collagen was present in higher amounts in cortical segments than core samples (72.8 ± 35.14 vs. 37.3 ± 16.99 µg/mgww, p = 0.0005). sGAG was also shown to be more prevalent in cortical segments (25.47 ± 11.59 vs. 12.17 ± 7.15 µg/mgww, p < 0.0001). The concentrations of collagen and sGAG demonstrated a positive correlation (R2 = 0.44, p = 0.0004). Collagen and sGAG content decreased with the age of the donor (p = 0.001 and p < 0.0001, respectively), but donor gender did not appear to affect collagen or sGAG content (p = 0.62 and p = 0.43, respectively). Conclusion: Collagen and sGAG content was higher in cortical segments of cadaveric rib cartilage than in core segments, and higher in samples from younger cadavers as well.

The survival of metallic residues from gunshot wounds in cremated bone: a SEM-EDX study.

The research and analysis of gunshot residues has a relevant role in the examination of gunshot wounds. Nevertheless, very little literature exists concerning gunshot wounds on charred material. In this study, 16 adult bovine ribs (eight still with soft tissues and eight totally skeletonized) underwent a shooting test with two types of projectiles (9 mm full metal-jacketed bullet and 9 mm unjacketed bullet). Each rib then underwent a charring process in an electric oven, reaching the stage of complete calcination at 800°C. The area of each entrance wound was analyzed before and after the carbonization process via a scanning electron microscope (SEM) equipped with an energy dispersive X-ray analyzer (EDX). In each sample, metallic residues composed of lead, barium, and antimony were found. These metallic residues were thus preserved also after exposure to the extremely high temperatures reached within the oven, especially with unjacketed bullets, although the particles seem to be more irregular in shape as a result of the heating process. In conclusion, this study proved that gunshot residues survive extremely high temperatures and can be detected via SEM/EDX even in cases of charred tissues.

The survival of metallic residues from gunshot wounds in cremated bone: a radiological study.

In literature, many studies have been performed in order to investigate the presence of GSR ("gunshot residue") and metallic residues in general with radiological techniques on several types of material, but the survival of metallic residues on charred samples has never been systematically performed. In this study, 31 adult bovine ribs underwent a shooting test. Every rib was shot with a single bullet, at a near-contact shooting distance, using two kinds of projectile: 17 samples were shot with a full metal-jacketed bullet and the remaining 14 with an unjacketed bullet. After the shooting test, every rib underwent a "charring cycle" in an electric oven up to 800°C. Every sample underwent radiological investigation with conventional radiography, before and after the burning process, to evaluate any changes in number and distribution of metallic residues. Radiographs showed survival of radiopaque residues in every sample, even after the charring process, especially when the bullet used was of the unjacketed type.

The men of Nelson's navy: a comparative stable isotope dietary study of late 18th century and early 19th century servicemen from Royal Naval Hospital burial grounds at Plymouth and Gosport, England.

We present stable isotopic analyses of collagen from 80 servicemen excavated from the late 18th/early 19th century naval hospitals at Plymouth (50) and Haslar, Gosport (30) in southern England. Historical records suggest that, the diets of these two populations should be essentially identical. While δ(15) N of the rib collagen confirmed that naval servicemen were relatively well-catered for in terms of meat allowance (Plymouth average δ(15) N = 11.1‰, Gosport = 11.9‰), stable carbon isotope analysis produced average values for the two assemblages, which were significantly different (Plymouth average δ(13) C = -18.8‰, Gosport = -20.0‰). We postulate that these differences stem from divergent naval postings, with a greater proportion of Plymouth individuals serving in areas that entailed a greater input of C(4) foodstuffs. By comparison with published data from approximately contemporary burials at Snake Hill, Ontario, Canada and Chesapeake Bay, Virginia, we suggest that this area is the east coast of North America. For 15 of the 30 individuals from Gosport, we have data on ribs, femur, and dentine from the same skeleton, which appear to show that they came from a variety of locations in their preadolescence, but converged in dietary terms onto a "naval average," which is consistent with historical evidence for recruitment patterns into the Navy at the time. By comparison with published data from skeletons recovered from the wreck of the Mary Rose (sank 1545), we conclude that this naval diet was virtually unchanged from the 16th century to the end of the 18th century.

Stable isotope evidence for sex- and status-based variations in diet and life history at medieval Trino Vercellese, Italy.

The medieval period in Europe was a time of unprecedented social complexity that affected human diet. The diets of certain subgroups-for example, children, women, and the poor-are chronically underrepresented in historical sources from the medieval period. To better understand diet and the distribution of foods during the medieval period, we investigated stable carbon and nitrogen isotope ratios of 30 individuals from Trino Vercellese, Northern Italy (8th-13th c.). Specifically, we examined diet differences between subgroups (males and females, and high- and low-status individuals), and diet change throughout the life course among these groups by comparing dentine and bone collagen. Our results show a diet based on terrestrial resources with input from C(4) plants, which could include proso and/or foxtail millet. Diets of low-status males differ from those of females (both status groups) and of high-status males. These differences develop in adulthood. Childhood diets are similar among the subgroups, but sex- and status-based differences appear in adulthood. We discuss the possibility of cultural buffering and dietary selectivity of females and high-status individuals.

A single change in the aptamer of the Lactiplantibacillus plantarum rib operon riboswitch severely impairs its regulatory activity and leads to a vitamin B2 - overproducing phenotype.

Manufacturing of probiotics and functional foods using lactic acid bacteria (LAB) that overproduce vitamin B2 has gained growing interest due to ariboflavinosis problems affecting populations of both developing and affluent countries. Two isogenic Lactiplantibacillus plantarum strains, namely a riboflavin-producing parental strain (UFG9) and a roseoflavin-resistant strain (B2) that carries a mutation in the FMN-aptamer of the potential rib operon riboswitch, were analysed for production and intra- and extracellular accumulation of flavins, as well as for regulation of the rib operon expression. Strain B2 accumulated in the medium one of the highest levels of riboflavin+FMN ever reported for LAB, exceeding by ~ 25 times those accumulated by UFG9. Inside the cells, concentration of FAD was similar in both strains, while that of riboflavin+FMN was ~ 8-fold higher in B2. Mutation B2 could decrease the stability of the aptamer's regulatory P1 helix even in the presence of the effector, thus promoting the antiterminator structure of the riboswitch ON state. Although the B2-mutant riboswitch showed an impaired regulatory activity, it retained partial functionality being still sensitive to the effector. The extraordinary capacity of strain B2 to produce riboflavin, together with its metabolic versatility and probiotic properties, can be exploited for manufacturing multifunctional foods.

Delayed osteoblastic differentiation and bone development in Cx43 knockout mice.

GJA1 gene (Connexin43, also known as Cx43) is the most abundant gap junction protein isoform in animal cells and is associated with bone development in embryos. The objective of the present work was to evaluate in vivo osteal development in GJA1-deficient fetal mice through determination of the histological and molecular alterations induced by partial or total deletion of the GJA1 gene. Heterozygous C57BL/6 mice (HT) harboring a null mutation of the GJA1 gene were mated, and pregnant females were submitted to euthanasia and Caesarean section from 12.5 to 19.5 days post coitum (dpc). HT (GJA1(+/-)) and homozygous (GJA1(-/- )) knockout (KO) mutants and wild-type (WT) fetuses were identified by polymerase chain reaction (PCR), and development curves were constructed on the basis of fetus weight and crown-rump length. Histopathological, histochemical, and real-time PCR analyses were performed in order to assess the expression of markers associated with bone development, namely, osteocalcin, osteopontin, alkaline phosphatase, RUNX2, GJA1, GJC1 (Cx45), and GJA3 (Cx46). HT and KO fetuses exhibited delays in the differentiation of osteoblasts and, consequently, in bone development in comparison with the WT group. Additionally, less deposition of mineralized and osteoid matrix was observed in GJA1-deficient fetuses. Bone development in KO fetuses was delayed through the moment of birth, but in HT animals the delay only extended until 17.5 dpc, following which development was normalized. The expression of genes coding for osteocalcin, osteopontin, alkaline phosphatise, and RUNX2 were also delayed in GJA1-deficient fetuses. Animals that exhibited a lower expression of GJA1 presented delayed expression of the GJC1 and GJA3 genes and their corresponding protein products in the bone tissue. The results of the present study contribute to our understanding of the function of GJA1 during bone development and suggest that GJC1 could play a role in restoring intercellular communication in GJA1-deficient mice.

Using stable isotope analysis to examine the effect of economic change on breastfeeding practices in Spitalfields, London, UK.

Breastfeeding patterns were subject to a number of fads in 18th and 19th century Britain. Feeding infants by hand, rather than maternal breastfeeding or wet-nursing, became more prevalent among both the wealthy and poor. Substitute foods may have been a convenient alternative for mothers employed away from the household. This study used stable isotope ratio analysis to examine the weaning schedule in the 18th and 19th century skeletal assemblage from Spitalfields, London, UK. Analysis of 72 juvenile ribs revealed δ(15) N elevations of 2-3‰ above the adult mean for individuals up to the age of two, while elevations of 1-2‰ were observed in δ(13) C for the first year of life. This suggests that the introduction of solid foods took place before the end of the first year, and that breastfeeding had entirely ceased by 2 years of age. The age at death of many of these infants is known from historical records, and can be used to pinpoint the amount of time required for the breast milk signal to be observed in the stable isotope ratios of rib collagen. Results show that a δ(15) N elevation can be detected in the ribs of individuals who died as young as 5-6 weeks. Not all individuals at Spitalfields were breastfed, and there may not have been a single uniformly practiced weaning scheme. There is, however, more evidence for prolonged breastfeeding during the 19th century than the 18th century.