Expression of the Metalloproteinase ADAM8 Is Upregulated in Liver Inflammation Models and Enhances Cytokine Release In Vitro

Acute and chronic liver inflammation is driven by cytokine and chemokine release from various cell types in the liver. Here, we report that the induction of inflammatory mediators is associated with a yet undescribed upregulation of the metalloproteinase ADAM8 in different murine hepatitis models. We further show the importance of ADAM8 expression for the production of inflammatory mediators in cultured liver cells. As a model of acute inflammation, we investigated liver tissue from lipopolysaccharide- (LPS-) treated mice in which ADAM8 expression was markedly upregulated compared to control mice. In vitro, stimulation with LPS enhanced ADAM8 expression in murine and human endothelial and hepatoma cell lines as well as in primary murine hepatocytes. The enhanced ADAM8 expression was associated with an upregulation of TNF-α and IL-6 expression and release. Inhibition studies indicate that the cytokine response of hepatoma cells to LPS depends on the activity of ADAM8 and that signalling by TNF-α can contribute to these ADAM8-dependent effects. The role of ADAM8 was further confirmed with primary hepatocytes from ADAM8 knockout mice in which TNF-α and IL-6 induction and release were considerably attenuated. As a model of chronic liver injury, we studied liver tissue from mice undergoing high-fat diet-induced steatohepatitis and again observed upregulation of ADAM8 mRNA expression compared to healthy controls. In vitro, ADAM8 expression was upregulated in hepatoma, endothelial, and stellate cell lines by various mediators of steatohepatitis including fatty acid (linoleic-oleic acid), IL-1β, TNF-α, IFN-γ, and TGF-β. Upregulation of ADAM8 was associated with the induction and release of proinflammatory cytokines (TNF-α and IL-6) and chemokines (CX3CL1). Finally, knockdown of ADAM8 expression in all tested cell types attenuated the release of these mediators. Thus, ADAM8 is upregulated in acute and chronic liver inflammation and is able to promote inflammation by enhancing expression and release of inflammatory mediators

Morphology and physiology of motor neurons innervating labral muscles of locusta migratoria

Major animal taxa, known as phyla, each represents a basic bauplan. Among the diversity of phyla, arthropods are the most dominant on the earth. The conspicuous feature of the arthropod bauplan is the grouping of body segments into specialized units by the phenomenon of tagmosis which in insect has given rise to three body regions; head, thorax and abdomen. But the number and the nature of segments involved in composition of the insect head is still a matter of dispute among today’s evolutionary biologists. From the beginning of the dispute, there was general agreement that three mouthparts segments (mandible, maxillae, and labium) take part in the head composition, and their ganglia fuse to form the suboesophageal ganglion of the insect nervous system. However, the nature of the segments anterior to the mandible (labral/ocular, antennal, and intercalary segments) remained controversial. Modern molecular, palaeontological, and neuroanatomical studies showed that the insect head is composed of six segments; occular, antennal, intercalary, and three gnathal segments. But conflicting views are still present regarding the nature and segmental affiliation of the insect labrum. Several such modern studies imply that the labrum is a highly modified appendage of the intercalary segment, and sensory innervation of the embryonic and the adult labrum shares many similarities with that of head and thoracic appendages. However, defining criterion of an appendage implies that every appendage must be innervated by its own segmental ganglion, and the innervation pattern must include both sensory and motor elements. The aim of thesis is to investigate the motor innervation pattern of muscles of the labrum in the adult locust to reveal the evolutionary changes underlying neuronal circuitry, which might help to decide whether the labrum represents a structure that has developed during the evolution by fusion of paired appendages associated with the intercalary segment. Using Neurobiotin as a retrograde neuronal tracer, specific motor nerves of individual labral muscles were stained. Results show that all labral muscles receive innervation from both the tritocerebral lobes of the brain and the suboesophageal ganglion, except for M1, the labral compressor muscle. M1 is innervated by two motor neurons. Both tritocerebral lobes contain one soma. The axons of both neurons branch in the periphery to innervate ipsi- and contralateral muscles. The labral anterior retractor muscles, M2, are innervated by 6 tritocerebral motor neurons, with 3 somata located in each tritocerebral lobe. Their axons cross the midline in a distinct commissure between the two muscles. The labral posterior retractor muscles, M3 and anterior dilator muscles of foregut, M38, are innervated from the ipsilateral tritocerebrum only. Both muscles together receive 8 motor axons and there are some common motor neurons that innervate both muscles. In addition, M2, M3 and M38 also receive common innervation from neurons located in the suboesophageal ganglion (SOG). In relation to the muscle, their somata are located on the ipsi- and the contralateral sides of the SOG. Their axons ascend through the circumoesophageal connectives and proceed into the frontal connectives without forming ramifications in the tritocerebral lobes. These axons also cross the midline in the periphery to reach muscles on both sides of the head. They either cross within the commissure between muscles M2, or through the frontal ganglion. Further more, electrophysiology of labral muscles in isolated insect heads was investigated by intracellular recordings from the fibers of individual muscles. The recordings showed that all postsynaptic potentials were excitatory and no inhibitory synaptic inputs to labral muscles were found during the present study. Similarly simultaneous intracellular recording from fibers of labral muscles and suboesophageal ganglion motor neurons also revealed no inhibitory synaptic inputs to labral muscles, and showed that suboesophageal neurons are common excitatory motor neurons. The innervation pattern was compared with that of the other body appendages of locust, and it was concluded that labral muscles are innervated from both sides of the CNS and their innervation pattern is bilaterally symmetric as that of other appendages. The participation of both tritocerebral and suboesophageal ganglion motor neurons supports the notion that the labrum is innervated by the so-called intercalary segment. The innervation pattern also supports the idea that the labrum derives from fused appendages: There are two symmetrical sets of motor neurons. Some of them cross the midline in the periphery. This indicates that during the evolution of the labrum its motor innervation pattern was reconfigured in the periphery, while the original bilateral sets of central neurons remained unchanged

Morphology and physiology of motor neurons innervating labral muscles of locusta migratoria

Major animal taxa, known as phyla, each represents a basic bauplan. Among the diversity of phyla, arthropods are the most dominant on the earth. The conspicuous feature of the arthropod bauplan is the grouping of body segments into specialized units by the phenomenon of tagmosis which in insect has given rise to three body regions; head, thorax and abdomen. But the number and the nature of segments involved in composition of the insect head is still a matter of dispute among today’s evolutionary biologists. From the beginning of the dispute, there was general agreement that three mouthparts segments (mandible, maxillae, and labium) take part in the head composition, and their ganglia fuse to form the suboesophageal ganglion of the insect nervous system. However, the nature of the segments anterior to the mandible (labral/ocular, antennal, and intercalary segments) remained controversial. Modern molecular, palaeontological, and neuroanatomical studies showed that the insect head is composed of six segments; occular, antennal, intercalary, and three gnathal segments. But conflicting views are still present regarding the nature and segmental affiliation of the insect labrum. Several such modern studies imply that the labrum is a highly modified appendage of the intercalary segment, and sensory innervation of the embryonic and the adult labrum shares many similarities with that of head and thoracic appendages. However, defining criterion of an appendage implies that every appendage must be innervated by its own segmental ganglion, and the innervation pattern must include both sensory and motor elements. The aim of thesis is to investigate the motor innervation pattern of muscles of the labrum in the adult locust to reveal the evolutionary changes underlying neuronal circuitry, which might help to decide whether the labrum represents a structure that has developed during the evolution by fusion of paired appendages associated with the intercalary segment. Using Neurobiotin as a retrograde neuronal tracer, specific motor nerves of individual labral muscles were stained. Results show that all labral muscles receive innervation from both the tritocerebral lobes of the brain and the suboesophageal ganglion, except for M1, the labral compressor muscle. M1 is innervated by two motor neurons. Both tritocerebral lobes contain one soma. The axons of both neurons branch in the periphery to innervate ipsi- and contralateral muscles. The labral anterior retractor muscles, M2, are innervated by 6 tritocerebral motor neurons, with 3 somata located in each tritocerebral lobe. Their axons cross the midline in a distinct commissure between the two muscles. The labral posterior retractor muscles, M3 and anterior dilator muscles of foregut, M38, are innervated from the ipsilateral tritocerebrum only. Both muscles together receive 8 motor axons and there are some common motor neurons that innervate both muscles. In addition, M2, M3 and M38 also receive common innervation from neurons located in the suboesophageal ganglion (SOG). In relation to the muscle, their somata are located on the ipsi- and the contralateral sides of the SOG. Their axons ascend through the circumoesophageal connectives and proceed into the frontal connectives without forming ramifications in the tritocerebral lobes. These axons also cross the midline in the periphery to reach muscles on both sides of the head. They either cross within the commissure between muscles M2, or through the frontal ganglion. Further more, electrophysiology of labral muscles in isolated insect heads was investigated by intracellular recordings from the fibers of individual muscles. The recordings showed that all postsynaptic potentials were excitatory and no inhibitory synaptic inputs to labral muscles were found during the present study. Similarly simultaneous intracellular recording from fibers of labral muscles and suboesophageal ganglion motor neurons also revealed no inhibitory synaptic inputs to labral muscles, and showed that suboesophageal neurons are common excitatory motor neurons. The innervation pattern was compared with that of the other body appendages of locust, and it was concluded that labral muscles are innervated from both sides of the CNS and their innervation pattern is bilaterally symmetric as that of other appendages. The participation of both tritocerebral and suboesophageal ganglion motor neurons supports the notion that the labrum is innervated by the so-called intercalary segment. The innervation pattern also supports the idea that the labrum derives from fused appendages: There are two symmetrical sets of motor neurons. Some of them cross the midline in the periphery. This indicates that during the evolution of the labrum its motor innervation pattern was reconfigured in the periphery, while the original bilateral sets of central neurons remained unchanged

Assemblage of pollinator communities in four widely isolated nature reserves of southern Punjab, Pakistan

Pollinators are key components of the global biodiversity, providing vital ecosystem services of pollination to crops and wild plants. Understanding pollinator assemblage and how it changes spatially is important in order to device effective ecosystem management planning. This is the first empirical study to explore pollinator community structure in four widely isolated nature reserves of southern Punjab, Pakistan, i.e., the central agricultural plain (Pirowal Forest), the Indus delta plain (Ghazi Ghat Wetland), hilly mountains (Fort Munro), and a sandy desert (Lal Suhanra Forest). Six assemblage parameters, i.e., abundance, richness, dominance, evenness, Simpson index, and Shannon-Wiener index, were studied over seven consecutive months (February to August 2012). Maximum abundance and richness of the pollinators were recorded at Ghazi Ghat (1354 individuals and 84 species), followed by Pirowal (1331 individuals and 80 species), Lal Suhanra (1197 individuals and 77 species), and Fort Munro (808 individuals and 65 species). The highest values of Simpson and evenness indexes were recorded at Fort Munro, while the Shannon-Wiener and dominance indexes were highest at Ghazi Ghat and Pirowal. Species richness was highest for bees, while it was lowest for flies, yet the latter were the highest in abundance among the four pollinator groups. The current study is a first account of pollinator assemblage structure in four widely isolated forest reserves of Punjab, Pakistan. This will support ecosystem management planning, and opens doors for further research in order to reveal complexities in pollinator assemblage structure especially in relation with plant communities. Keywords: Pollinators, Assemblage, Forests, Punjab, Pakista

Expression of the Metalloproteinase ADAM8 Is Upregulated in Liver Inflammation Models and Enhances Cytokine Release In Vitro

Acute and chronic liver inflammation is driven by cytokine and chemokine release from various cell types in the liver. Here, we report that the induction of inflammatory mediators is associated with a yet undescribed upregulation of the metalloproteinase ADAM8 in different murine hepatitis models. We further show the importance of ADAM8 expression for the production of inflammatory mediators in cultured liver cells. As a model of acute inflammation, we investigated liver tissue from lipopolysaccharide- (LPS-) treated mice in which ADAM8 expression was markedly upregulated compared to control mice. In vitro, stimulation with LPS enhanced ADAM8 expression in murine and human endothelial and hepatoma cell lines as well as in primary murine hepatocytes. The enhanced ADAM8 expression was associated with an upregulation of TNF-α and IL-6 expression and release. Inhibition studies indicate that the cytokine response of hepatoma cells to LPS depends on the activity of ADAM8 and that signalling by TNF-α can contribute to these ADAM8-dependent effects. The role of ADAM8 was further confirmed with primary hepatocytes from ADAM8 knockout mice in which TNF-α and IL-6 induction and release were considerably attenuated. As a model of chronic liver injury, we studied liver tissue from mice undergoing high-fat diet-induced steatohepatitis and again observed upregulation of ADAM8 mRNA expression compared to healthy controls. In vitro, ADAM8 expression was upregulated in hepatoma, endothelial, and stellate cell lines by various mediators of steatohepatitis including fatty acid (linoleic-oleic acid), IL-1β, TNF-α, IFN-γ, and TGF-β. Upregulation of ADAM8 was associated with the induction and release of proinflammatory cytokines (TNF-α and IL-6) and chemokines (CX3CL1). Finally, knockdown of ADAM8 expression in all tested cell types attenuated the release of these mediators. Thus, ADAM8 is upregulated in acute and chronic liver inflammation and is able to promote inflammation by enhancing expression and release of inflammatory mediators

Role of pollination in yield and physicochemical properties of tomatoes (Lycopersicon esculentum)

Very little is known about pollination and its effects on the yield and physicochemical properties of flowering plants in tropical countries. Wind and insect pollinators are among our natural resources because pollination is the most important ecosystem service performed by wind and insects, and is vital to the socio-economic status of human beings. In this experiment, different pollination methods for tomato plants were examined. Self-pollination was encouraged by covering the plants with a plastic sheet. Wind and insects were excluded from these plants, and thus only self-pollination was possible. The experiment occurred during the flowering stage. Wind-pollinated plants were covered with a muslin cloth, which excluded insects, and only wind could pass through the cloth. For insect pollination, plants remained uncovered, allowing free access to insects to pollinate the flowers. At fruit maturity, when fruits were completely red, fruits from each treatment were harvested on the same date and under the same conditions. Results illustrated the substantial importance of insects as pollinators of tomato crops. Open field had greater tomato yield and positive effects on physicochemical properties on fruit than under self and wind pollination. Keywords: Pollination methods, Tomato, Lycopersicon esculentum, Physicochemical properties, Yiel

Global fertility in 204 countries and territories, 1950–2021, with forecasts to 2100: a comprehensive demographic analysis for the Global Burden of Disease Study 2021

BackgroundAccurate assessments of current and future fertility—including overall trends and changing population age structures across countries and regions—are essential to help plan for the profound social, economic, environmental, and geopolitical challenges that these changes will bring. Estimates and projections of fertility are necessary to inform policies involving resource and health-care needs, labour supply, education, gender equality, and family planning and support. The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 produced up-to-date and comprehensive demographic assessments of key fertility indicators at global, regional, and national levels from 1950 to 2021 and forecast fertility metrics to 2100 based on a reference scenario and key policy-dependent alternative scenarios. MethodsTo estimate fertility indicators from 1950 to 2021, mixed-effects regression models and spatiotemporal Gaussian process regression were used to synthesise data from 8709 country-years of vital and sample registrations, 1455 surveys and censuses, and 150 other sources, and to generate age-specific fertility rates (ASFRs) for 5-year age groups from age 10 years to 54 years. ASFRs were summed across age groups to produce estimates of total fertility rate (TFR). Livebirths were calculated by multiplying ASFR and age-specific female population, then summing across ages 10–54 years. To forecast future fertility up to 2100, our Institute for Health Metrics and Evaluation (IHME) forecasting model was based on projections of completed cohort fertility at age 50 years (CCF50; the average number of children born over time to females from a specified birth cohort), which yields more stable and accurate measures of fertility than directly modelling TFR. CCF50 was modelled using an ensemble approach in which three sub-models (with two, three, and four covariates variously consisting of female educational attainment, contraceptive met need, population density in habitable areas, and under-5 mortality) were given equal weights, and analyses were conducted utilising the MR-BRT (meta-regression—Bayesian, regularised, trimmed) tool. To capture time-series trends in CCF50 not explained by these covariates, we used a first-order autoregressive model on the residual term. CCF50 as a proportion of each 5-year ASFR was predicted using a linear mixed-effects model with fixed-effects covariates (female educational attainment and contraceptive met need) and random intercepts for geographical regions. Projected TFRs were then computed for each calendar year as the sum of single-year ASFRs across age groups. The reference forecast is our estimate of the most likely fertility future given the model, past fertility, forecasts of covariates, and historical relationships between covariates and fertility. We additionally produced forecasts for multiple alternative scenarios in each location: the UN Sustainable Development Goal (SDG) for education is achieved by 2030; the contraceptive met need SDG is achieved by 2030; pro-natal policies are enacted to create supportive environments for those who give birth; and the previous three scenarios combined. Uncertainty from past data inputs and model estimation was propagated throughout analyses by taking 1000 draws for past and present fertility estimates and 500 draws for future forecasts from the estimated distribution for each metric, with 95% uncertainty intervals (UIs) given as the 2·5 and 97·5 percentiles of the draws. To evaluate the forecasting performance of our model and others, we computed skill values—a metric assessing gain in forecasting accuracy—by comparing predicted versus observed ASFRs from the past 15 years (2007–21). A positive skill metric indicates that the model being evaluated performs better than the baseline model (here, a simplified model holding 2007 values constant in the future), and a negative metric indicates that the evaluated model performs worse than baseline. FindingsDuring the period from 1950 to 2021, global TFR more than halved, from 4·84 (95% UI 4·63–5·06) to 2·23 (2·09–2·38). Global annual livebirths peaked in 2016 at 142 million (95% UI 137–147), declining to 129 million (121–138) in 2021. Fertility rates declined in all countries and territories since 1950, with TFR remaining above 2·1—canonically considered replacement-level fertility—in 94 (46·1%) countries and territories in 2021. This included 44 of 46 countries in sub-Saharan Africa, which was the super-region with the largest share of livebirths in 2021 (29·2% [28·7–29·6]). 47 countries and territories in which lowest estimated fertility between 1950 and 2021 was below replacement experienced one or more subsequent years with higher fertility; only three of these locations rebounded above replacement levels. Future fertility rates were projected to continue to decline worldwide, reaching a global TFR of 1·83 (1·59–2·08) in 2050 and 1·59 (1·25–1·96) in 2100 under the reference scenario. The number of countries and territories with fertility rates remaining above replacement was forecast to be 49 (24·0%) in 2050 and only six (2·9%) in 2100, with three of these six countries included in the 2021 World Bank-defined low-income group, all located in the GBD super-region of sub-Saharan Africa. The proportion of livebirths occurring in sub-Saharan Africa was forecast to increase to more than half of the world's livebirths in 2100, to 41·3% (39·6–43·1) in 2050 and 54·3% (47·1–59·5) in 2100. The share of livebirths was projected to decline between 2021 and 2100 in most of the six other super-regions—decreasing, for example, in south Asia from 24·8% (23·7–25·8) in 2021 to 16·7% (14·3–19·1) in 2050 and 7·1% (4·4–10·1) in 2100—but was forecast to increase modestly in the north Africa and Middle East and high-income super-regions. Forecast estimates for the alternative combined scenario suggest that meeting SDG targets for education and contraceptive met need, as well as implementing pro-natal policies, would result in global TFRs of 1·65 (1·40–1·92) in 2050 and 1·62 (1·35–1·95) in 2100. The forecasting skill metric values for the IHME model were positive across all age groups, indicating that the model is better than the constant prediction. InterpretationFertility is declining globally, with rates in more than half of all countries and territories in 2021 below replacement level. Trends since 2000 show considerable heterogeneity in the steepness of declines, and only a small number of countries experienced even a slight fertility rebound after their lowest observed rate, with none reaching replacement level. Additionally, the distribution of livebirths across the globe is shifting, with a greater proportion occurring in the lowest-income countries. Future fertility rates will continue to decline worldwide and will remain low even under successful implementation of pro-natal policies. These changes will have far-reaching economic and societal consequences due to ageing populations and declining workforces in higher-income countries, combined with an increasing share of livebirths among the already poorest regions of the world. FundingBill & Melinda Gates Foundation