Trastuzumab activates allogeneic or autologous antibody-dependent cellular cytotoxicity against malignant rhabdoid tumor cells and interleukin-2 augments the cytotoxicity.

PURPOSE: Malignant rhabdoid tumor (MRT) is an early childhood cancer with poor prognosis. Trastuzumab, a humanized monoclonal antibody against human epidermal growth factor receptor-2 (HER-2), has been shown to be effective against breast cancer and other cancers. We investigated the effect of trastuzumab on MRT cell lines. EXPERIMENTAL DESIGN: We examined expression of HER-2 on four MRT cell lines and two tumor tissues by indirect immunofluorescence, flow cytometry, and immunohistochemistry. The effect of trastuzumab against MRT cells was examined by cell growth assay. To observe the antibody-dependent cellular cytotoxicity of effector cells, we examined the cytotoxicity of trastuzumab in combination with allogeneic or autologous human peripheral blood mononuclear cells with and without IL-2 using the chromium release assay. RESULTS: All four MRT cell lines and both MRT tissues expressed HER-2 protein. Trastuzumab alone did not reduce the viability of the MRT cell lines. On the other hand, the cytotoxicity of trastuzumab against each of the MRT cell lines was significantly increased by the presence of allogeneic and autologous peripheral blood mononuclear cells (P < 0.01). There was a strong correlation coefficient (r = 0.825) between HER-2 expression and the cytotoxicity enhanced by trastuzumab. Moreover, trastuzumab in combination with peripheral blood mononuclear cells augmented by interleukin-2 (IL-2) was significantly more cytotoxic than trastuzumab alone or IL-2 alone (P < 0.01). CONCLUSIONS: Our results indicate that (1) trastuzumab can exert antitumor effects on MRT cells by using the antibody-dependent cellular cytotoxicity of effector cells and (2) IL-2 can enhance the cytotoxicity of trastuzumab against MRT cells.     

Enhancement of adipogenesis induction by conditioned media obtained from cancer cells

White adipose tissue is a multifunctional endocrine organ that synthesizes and secretes cytokine-like proteins termed adipokines. In the present study, the effects of cancer-derived medium on adipogenesis were examined. We prepared conditioned media from cancer cell lines, and cultured preadipocytes in the conditioned media. After 10 days of culture, intracellular lipid droplet accumulation was measured. We observed that the conditioned media significantly induced adipogenesis or enhanced the adipogenesis induced by adipogenesis inducers. Although we could not define the factors, these undefined factors derived from cancer cells may induce adipogenesis, and the resulting adipogenesis may affect cancer development.     

An autopsy case of Alzheimer's disease presenting with primary progressive aphasia: A clinicopathological and immunohistochemical study

This report describes an autopsied Alzheimer's disease (AD) patient with primary progressive aphasia (PPA) as an early symptom. The patient developed a progressive speech disturbance at the age of 70 years, and difficulty in comprehension became apparent 2 years later. Magnetic resonance imaging scan disclosed asymmetrical brain atrophy, predominantly on the left temporal lobe. At the age of 74 years, the patient's dementia rapidly progressed with parkinsonism and he died after a disease duration of 6 years. At autopsy, the brain showed a marked temporo‐frontal lobe atrophy, predominantly on the left side. There was severe neuronal loss with gliosis and tissue rarefaction in the atrophied cerebral cortex and amygdala. Many neurofibrillary tangles with neuropil threads were found in the cerebral cortex. Numerous amyloid deposits were distributed throughout the cerebral cortex, accompanied by amyloid angiopathies. This patient was clinically diagnosed with temporal lobe‐dominant Pick's disease, although the possibility of corticobasal degeneration was made. The neuropathological diagnosis was AD with asymmetrical brain atrophy and widespread amyloid angiopathies.     

Anthropogenic impacts on Late Holocene land-cover change and floristic biodiversity loss in tropical southeastern Asia

Southern China and Southeast Asia witnessed some of their most significant economic and social changes relevant to human land use during the Late Holocene, including the intensification and spread of rice agriculture. Despite rice growth being associated with a number of earth systems impacts, how these changes transformed tropical vegetation in this region of immense endemic biodiversity remains poorly understood. Here, we compile a pollen dataset incorporating ∼150,000 identifications and 233 pollen taxa to examine past changes in floral biodiversity, together with a compilation of records of forest decline across the region using 14 pollen records spanning lowland to mountain sites. Our results demonstrate that the rise of intensive rice agriculture from approximately 2,000 y ago led not only to extensive deforestation but also to remarkable changes of vegetation composition and a reduction in arboreal diversity. Focusing specifically on the Tertiary relic tree species, the freshwater wetland conifer Glyptostrobus (Glyptostrobus pensilis), we demonstrate how key species that had survived changing environmental conditions across millions of years shrank in the face of paddy rice farming and human disturbance.

The tropical forests of southern China and Southeast Asia (which we term together here as southeastern Asia) are renowned for their rich and unique endemic flora that, in many cases, can trace its origins back to the end of the Mesozoic approximately 65 million y ago (1). The relatively stable geological conditions of the region since the Late Cretaceous period have meant that it has frequently been seen as a long-term refugium for relict plant species, and is home to one of the highest concentrations of endemism around the globe (2, 3) and at least 4 of the 25 globally important biodiversity hotspots (4). Today, however, the tropical forests of southeastern Asia are among the most threatened land-based ecosystems on the planet, witnessing some of the fastest rates of deforestation in the face of urban population growth, changes in land-use, and human-induced climate change since the 20th century (2, 5). Agricultural expansion is seen as one of the major drivers of forest loss in southeastern Asia, with the conversion of tropical forest ecosystems to plantations or subsistence fields leading to drastic reductions in biodiversity (6–9).While the anthropogenically induced changes of these supposedly “ancient” ecosystems are often considered recent, there is a growing consideration that humans have interacted with tropical vegetation, including in southeastern Asia, across millennia, leaving legacies of biodiversity change, habitat manipulation, and forest cover that have lasting implications for economies, environments, and conservation policies today (10, 11). The arrival of rice cultivation in southern China and Southeast Asia took place around 4,000 y ago (ka), and the expansion of large-scale intensive rice agriculture in this area can be traced back to as early as 2.5 to 2 ka (12, 13), as the formation of coastal plains under freshwater conditions created expansive areas suitable for rice cultivation, together with a massive influx of rice farming immigrants and advances in agricultural technology, with potentially early impacts on tropical forest cover in the region. There have also been suggestions that the expansion of rice paddy fields, alongside domesticated water buffalo, in southeastern Asia may have resulted in a rise in methane emissions that fundamentally altered regional, and perhaps global, climate (14). Nevertheless, to test the degree to which these early agricultural expansions impacted tropical forest cover and biodiversity, and their relevance to ecosystem management in the 21st century, we require direct evidence for paleoecological change at the local and regional scale.Fossil pollen data from sediment cores can provide continuous records of past vegetation change and are widely used as a reliable proxy for forest-cover change (15, 16). Moreover, pollen data contain information on plant diversity and can thus also be used as a measure for past taxonomic richness. Although pollen records can struggle to capture the real plant diversity in tropical areas, where entomophilous taxa produce very few pollen grains and are thus usually underestimated in pollen assemblages, it is still possible to get meaningful results from pollen data. Supporting evidence comes in the form of the distinct latitudinal trend of palynological richness seen among woody taxa (17), indicating that the high diversity of vegetation characteristic of tropical regions can be revealed by pollen records. In addition, modern pollen studies undertaken in eastern Asia, including within both subtropical and tropical areas, suggest that modern pollen “rain” can successfully predict the natural biome, revealing a good relationship between palynological data and plant composition (18). The latest pollen productivities study in Hainan Island (19), which presents by far the most detailed pollen–vegetation survey from the northern part of tropical southeastern Asia, indicates that, at a regional scale—although there were differences between the modern pollen assemblages and related vegetation composition—the pollen types identified in surface soil samples captured the majority of investigated plant types (∼65%) at the family level. Moreover, this modern survey suggests that pollen assemblages of cultivated lands can be distinguished from recovered secondary woodlands, indicating that information of agriculture-induced vegetation change can be captured in pollen records in this part of the tropics (19).Here, we synthetically analyze pollen records in coastal areas of southeastern Asia that host northern tropical and southern subtropical moist broadleaf forests today, examining land-cover and floristic biodiversity change during the past 6,000 y. Our study time periods, spanning the last 6,000 y, are designed so as to focus on the known archaeological periods when the Neolithic subsistence economy mainly relied on hunting and gathering (before 3 ka) and when large-scale intensive rice agriculture began to emerge and expand (around 2.5 ka). First, 14 pollen records spanning lowland to mountain sites were systematically interrogated in order to reveal a comprehensive deforestation history for the study region. Then, we compiled a pollen dataset incorporating ∼150,000 identifications and 233 harmonized pollen morphological types from 9 pollen records to examine past changes in floral biodiversity. Furthermore, we examined how early paddy rice farming interacted with the now-endangered swamp conifer, Glyptostrobus pensilis, that had stood tall in the tropical forests of the region since the Tertiary period. Our data provide an example as to how regional paleoecological analysis can provide broader insights into the degree to which human economies and societies altered past landscapes and biota in the tropics, with potential ramifications for understanding the remnant vegetation and habitat distributions of the 21st century and the ongoing threats facing tropical forests in the “Anthropocene” (20).     

Efficient computation of Faith's phylogenetic diversity with applications in characterizing microbiomes

The number of publicly available microbiome samples is continually growing. As data set size increases, bottlenecks arise in standard analytical pipelines. Faith''s phylogenetic diversity (Faith''s PD) is a highly utilized phylogenetic alpha diversity metric that has thus far failed to effectively scale to trees with millions of vertices. Stacked Faith''s phylogenetic diversity (SFPhD) enables calculation of this widely adopted diversity metric at a much larger scale by implementing a computationally efficient algorithm. The algorithm reduces the amount of computational resources required, resulting in more accessible software with a reduced carbon footprint, as compared to previous approaches. The new algorithm produces identical results to the previous method. We further demonstrate that the phylogenetic aspect of Faith''s PD provides increased power in detecting diversity differences between younger and older populations in the FINRISK study''s metagenomic data.

In microbiome research, particular attention is given to evaluating the diversity of microbes within samples (The Human Microbiome Project Consortium 2012; Thompson et al. 2017; McDonald et al. 2018a). Alpha diversity (within sample diversity) represents a family of summary statistics that can summarize the breadth of diversity present in an environment. More recently, many examples have been reported on the associations between various host or environmental factors and alpha diversity of microbiomes, including country and diet in human guts (McDonald et al. 2018a), disease status in humans and canines (Gevers et al. 2014; Vázquez-Baeza et al. 2016), and the pH (Lauber et al. 2009), salinity (Thompson et al. 2017), and temperature (Zhou et al. 2016) of soils, among many others (Jeffery et al. 2016; Youngblut et al. 2019). A popular metric that accounts for the phylogenetic relatedness of the community members, Faith''s phylogenetic diversity (Faith''s PD) (Faith 1992), has been noted to be more sensitive in distinguishing disease factors in the human digestive system, relative to other alpha diversity indices (Scherson and Faith 2018; Youngblut et al. 2021).Modern DNA sequencing instruments have enabled microbiome studies at the scale of tens of thousands of samples, which presents a computational challenge for metrics that rely on a phylogeny, such as Faith''s PD. This metric is computed by summing the branch lengths (edge weights) of the phylogeny that exclusively represents the sequences contained in a biological sample. The amount of memory and number of necessary operations needed to calculate Faith''s PD depends on the number of edges in the phylogenetic tree, as well as the number of samples in the underlying data table.In today''s increasingly large and sparse data sets and meta-analyses, these phylogenetic trees and tables can exceed hundreds of thousands of samples and millions of tree tips (McDonald et al. 2018b). Recent advances have enabled efficient computation of the UniFrac metric for beta diversity. UniFrac is also a metric computed over phylogenetic trees (Lozupone and Knight 2005) and is mathematically related to Faith''s PD (Faith et al. 2009). Specifically, Striped UniFrac (McDonald et al. 2018b) improves upon previous UniFrac implementations (Hamady et al. 2010) by using space- and time-efficient tree data structures (Cordova and Navarro 2016) and reducing the number of vectors required to store intermediate scores in the tree.Additionally, the usefulness of techniques like Faith''s PD and UniFrac remains underexplored for metagenomics sequencing. Recent molecular protocol optimizations, such as SHOGUN (Hillmann et al. 2018), have enabled the metagenomic characterization of large human cohorts (Borodulin et al. 2015; Kaplan et al. 2019; Salosensaari et al. 2021). In this context, the applicability of Faith''s PD has largely been limited by the technical difficulties associated with constructing phylogenies from metagenomic features (Zhu et al. 2019). Efforts like the Web of Life (WoL) (Zhu et al. 2019) and Genome Taxonomy Database (GTDB) (Parks et al. 2018, 2020) are now addressing this issue by providing a phylogenomic tree as part of their database releases that can be used for phylogeny-informed analysis.Motivated by these advances in algorithms and resources for analyzing phylogenies, phylogenomic trees, and sparse data, we developed a new algorithm and implementation, stacked Faith''s phylogenetic diversity (SFPhD), for rapidly computing Faith''s PD. Additionally, we aim to demonstrate concrete benefits of phylogeny-informed analysis in metagenomic studies where this metric is less frequently used.     

Severe hypertension in infant with unilateral hypoplastic kidney

Unilateral renal hypoplasia unlike Ask-Upmark kidney (segmental renal hypoplasia) seldom causes hypertension. A case of an infant whose hypertension disappeared after removal of a hypoplastic kidney is reported. The removed kidney revealed no sign of Ask-Upmark lesion.     

Exercising the Sanger Sequencing Strategy for Variants Screening and Full-Length Genome of SARS-CoV-2 Virus during Alpha,Delta, and Omicron Outbreaks in Hiroshima

This study aimed to exercise the Sanger sequencing strategy for screening of variants among confirmed COVID-19 cases and validate our strategy against NGS strains in Hiroshima retrieved from GISAID. A total of 660 samples from confirmed COVID-19 cases underwent screening for variants by Sanger-based partial sequencing to the targeted spike gene (nt22,735~nt23,532) using an in-house-developed primer set. The identification of variants was done by unique checkpoints of base nucleotide changes in the targeted spike gene. Moreover, we amplified one full-length genome using Sanger method and an in-house-developed primer library. Using NGS strains of the same sampling period from GISAID, a phylogenetic tree was constructed to examine the distribution pattern of variants in Hiroshima and to validate our Sanger method. The modified primer set provided 100% validation and 99.2% amplification. PANGO Lineage R.1 was detected in late in the third wave, followed by Alpha (B.1.1.7) domination in the fourth wave, Delta (B.1.617.2) domination in the fifth wave, and Omicron (B.1.1.529) domination in the sixth wave, and there was no significant difference in viral copies between variants (p = 0.09). The variants showed different transmission patterns, but the distribution of variants is consistent to that shown by the phylogenetic tree. The Sanger method also provided successful amplification of the full-length genome of the SARS-CoV-2 virus. Our Sanger sequencing strategy was useful for the screening of SASR-CoV-2 variants without the need for full-genome amplification. The modified primer set was validated to use universally, which allows an understanding of the variants’ distribution in real time and provides the evidence for policy-making and the formulation or modification of preventive strategies.     

Association of serum fetuin-A with carotid arterial stiffness

OBJECTIVE: Fetuin-A is a circulating glycoprotein which is well characterized as an inhibitor of ectopic calcification. Vascular calcification commonly found in chronic kidney disease (CKD) patients is a predictor of cardiovascular death. Recently, several groups have demonstrated that low fetuin-A levels are associated with mortality in uraemic patients, possibly through regulation of vascular calcification. However, the physiological significance of fetuin-A in atherosclerosis remains unknown, except in specific conditions, such as vascular calcification in CKD patients. The objective of this study was to investigate the association between serum fetuin-A levels and arterial stiffness, a functional property of atherosclerosis, in healthy subjects. PATIENTS AND MEASUREMENTS: The study subjects comprised 141 healthy subjects. We measured serum fetuin-A levels and stiffness parameter beta for the common carotid artery, which was assessed by ultrasound using a phase-locked echo-tracking system. RESULTS: Simple regression analyses indicated that serum fetuin-A levels were significantly correlated with stiffness parameter beta (r = 0.200, P = 0.018). Multiple regression analyses showed that, besides age, fetuin-A (beta = 0.166, P = 0.033) independently contribute to the stiffness parameter beta (R(2) = 0.310, P < 0.0001). CONCLUSIONS: Serum fetuin-A level is associated with carotid arterial stiffness, independent of known atherogenic factors in healthy subjects.