Citizens can help to map putative transmission sites for snail-borne diseases.

INTRODUCTION: Schistosomiasis and fasciolosis are snail-borne diseases of great medical and veterinary health importance. The World Health Organization recommends complementing drug treatment with snail control and community involvement for disease elimination, but there is a general lack of snail experts and hence snail distribution data. Therefore, we adopted a citizen science approach and involved citizens in the monitoring of medically and veterinary important snail taxa. MATERIALS AND METHODS: Snail data was collected weekly by 25 trained citizen scientists (CSs) at 76 sites around southern Lake Albert (Uganda) for 20 months. At each site, snails were searched for 30 minutes, sorted, target snail hosts identified to genus level, counted and data submitted through a smartphone application. The quality of this data was assessed by comparing it to monthly data collected by an 'expert' malacologist using the same sampling protocol. Generalised binomial logistic and linear mixed-effects models were used to analyse the variables for agreement between the CSs and expert. FINDINGS: The binary agreement in presence/absence of Biomphalaria, Bulinus and Radix snails reported by the expert and CSs ranged between 70% and 86% (900 reports) with an average of 17% false negatives (sites wrongly defined as snail-free). The agreement for Biomphalaria and Radix increased with snail abundance, and false negatives decreased when the number of snails collected by citizens was aggregated per month. Site type significantly predicted binary agreement, which was lowest at lake sites (55%) and highest at spring sites (99%) with variations across genera. Similar temporal trends in snail abundance were recorded despite the expert reporting higher abundance. However, the relative abundance was consistent across site types. The match between the sites with highest Biomphalaria spp. abundance identified by CSs and expert was consistently high (~84.1%) and increased over time. CONCLUSIONS AND RECOMMENDATIONS: Our results demonstrate the potential of citizen science to map putative schistosomiasis transmission sites. We therefore argue that this inclusive, powerful and cost-effective approach can be more sustainable than top-down monitoring and intervention campaigns.

A machine learning approach for modeling the occurrence of the major intermediate hosts for schistosomiasis in East Africa.

Schistosomiasis, a prevalent water-borne disease second only to malaria, significantly impacts impoverished rural communities, primarily in Sub-Saharan Africa where over 90% of the severely affected population resides. The disease, majorly caused by Schistosoma mansoni and S. haematobium parasites, relies on freshwater snails, specifically Biomphalaria and Bulinus species, as crucial intermediate host (IH) snails. Targeted snail control is advisable, however, there is still limited knowledge about the community structure of the two genera especially in East Africa. Utilizing a machine learning approach, we employed random forest to identify key features influencing the distribution of both IH snails in this region. Our results reveal geography and climate as primary factors for Biomphalaria, while Bulinus occurrence is additionally influenced by soil clay content and nitrogen concentration. Favorable climate conditions indicate a high prevalence of IHs in East Africa, while the intricate connection with geography might signify either dispersal limitations or environmental filtering. Predicted probabilities demonstrate non-linear patterns, with Bulinus being more likely to occur than Biomphalaria in the region. This study provides foundational framework insights for targeted schistosomiasis prevention and control strategies in the region, assisting health workers and policymakers in their efforts.

Exploring the interplay between climate change and schistosomiasis transmission dynamics.

Schistosomiasis, a neglected tropical disease caused by parasitic worms, poses a major public health challenge in economically disadvantaged regions, especially in Sub-Saharan Africa. Climate factors, such as temperature and rainfall patterns, play a crucial role in the transmission dynamics of the disease. This study presents a deterministic model that aims to evaluate the temporal and seasonal transmission dynamics of schistosomiasis by examining the influence of temperature and rainfall over time. Equilibrium states are examined to ascertain their existence and stability employing the center manifold theory, while the basic reproduction number is calculated using the next-generation technique. To validate the model's applicability, demographic and climatological data from Uganda, Kenya, and Tanzania, which are endemic East African countries situated in the tropical region, are utilized as a case study region. The findings of this study provide evidence that the transmission of schistosomiasis in human populations is significantly influenced by seasonal and monthly variations, with incidence rates varying across countries depending on the frequency of temperature and rainfall. Consequently, the region is marked by both schistosomiasis emergencies and re-emergences. Specifically, it is observed that monthly mean temperatures within the range of 22-27 °C create favorable conditions for the development of schistosomiasis and have a positive impact on the reproduction numbers. On the other hand, monthly maximum temperatures ranging from 27 to 33 °C have an adverse effect on transmission. Furthermore, through sensitivity analysis, it is projected that by the year 2050, factors such as the recruitment rate of snails, the presence of parasite egg-containing stools, and the rate of miracidia shedding per parasite egg will contribute significantly to the occurrence and control of schistosomiasis infections. This study highlights the significant influence of seasonal and monthly variations, driven by temperature and rainfall patterns, on the transmission dynamics of schistosomiasis. These findings underscore the importance of considering climate factors in the control and prevention strategies of schistosomiasis. Additionally, the projected impact of various factors on schistosomiasis infections by 2050 emphasizes the need for proactive measures to mitigate the disease's impact on vulnerable populations. Overall, this research provides valuable insights to anticipate future challenges and devise adaptive measures to address schistosomiasis transmission patterns.

An accident waiting to happen? Exposing the potential of urogenital schistosomiasis transmission in the Lake Albert region, Uganda.

BACKGROUND: Urogenital schistosomiasis caused by the parasitic blood fluke Schistosoma haematobium is the most common form of that constitutes a majority of over 240 million schistosomiasis cases. The enigmatic absence of urogenital schistosomiasis in Uganda has, until now, been attributed to the absence of substantial populations of suitable snail intermediate hosts. METHODS: Malacological surveys were carried out in 73 sites southeast of Lake Albert, Uganda in October and November 2020. Collected snails were transported to the laboratory for identification. The snails were identified using partial mitochondrial cytochrome c oxidase subunit one and nuclear internal transcribed spacer barcoding. Schistosome infections in snails were also assessed using cercarial shedding and rapid diagnostic PCR techniques. RESULTS: We found Bulinus globosus and Bulinus nasutus productus, the main intermediate species in the transmission of S. haematobium in mainland East Africa. In this survey, B. globosus was more common than B. nasutus productus, with the former reported at four sites (total count = 188) and the latter reported at one site (total count = 79). Molecular testing revealed a high prevalence of Schistosoma bovis in B. nasutus productus (16%), but no S. haematobium infections were found. CONCLUSIONS: Given the abundance of snail hosts and the risky human water contact behaviours observed, we highlight the potential for urogenital schistosomiasis transmission in the region.

Historical DNA solves century-old mystery on sessility in freshwater gastropods.

Extinction rates are increasing unabatedly but resources available for conservation action are limited. Therefore, some conservationists are pushing for ecology- and evolution-based conservation choices, prioritizing taxa with phylogenetic and trait-based originality. Extinction of original taxa may result in a disproportionate loss of evolutionary innovations and potentially prevent transformative changes in living systems. Here, we generated historical DNA data from an almost 120-year-old syntype of the enigmatic sessile snail Helicostoa sinensis from the Three Gorges region of the Yangtze River (PR China), using a next-generation sequencing protocol developed for ancient DNA. In a broader phylogenetic context, we assessed the phylogenetic and trait-based originality of this enigmatic taxon to solve the century-old puzzle of sessility in freshwater gastropods. Our multi-locus data confirm the phylogenetic and trait-based originality of H. sinensis. It is an ultra-rare, subfamily-level taxon (Helicostoinae stat. nov.) within the family Bithyniidae, which exhibits the evolutionary innovation of sessility. While we conservatively classify H. sinensis as "Critically Endangered", there is mounting evidence of the biological annihilation of this endemic species. Although rapidly rising extinction rates in invertebrates are increasingly recognized, the potential loss of originality in these "little things that run the world" has received little attention. We therefore call for comprehensive surveys of originality in invertebrates, particularly from extreme environments such as rapids of large rivers, as a basis for urgently needed ecology- and evolution-based conservation decisions.

Long-term Assessment of Subchondral Bone Changes and Intralesional Bony Overgrowth After Third-Generation Autologous Chondrocyte Implantation: A Retrospective Study.

BACKGROUND: There is an increasing interest in subchondral bone changes and intralesional bony overgrowth (ILBO) after cartilage repair. Their clinical and predictive relevance is unclear and debated. PURPOSE: To evaluate the long-term development of ILBO and bone marrow edema-like signals (BMELSs) after autologous chondrocyte implantation (ACI) treatment of cartilage defects to find any predictive factors for their appearance. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: A total of 130 patients with 160 cartilage defects in the knee joint treated with third-generation ACI were included in this study. Radiological scores as the MOCART (magnetic resonance observation of cartilage repair tissue), the MOCART 2.0 and the 3D-MOCART using magnetic resonance imaging (MRI), and patient-reported outcome measures such as the Knee injury and Osteoarthritis Outcome Score (KOOS), International Knee Documentation Committee (IKDC) score, Noyes Sports Activity Rating Scale (NSARS) score, and Tegner Activity Scale (TAS) score were evaluated between 60 and 120 months (mean, 88 months) postoperatively. The radiological evaluation focused on the occurrence and size of subchondral bone changes, BMELSs, and ILBO during short-, medium-, and long-term follow-up. RESULTS: In long-term evaluation of clinical data, the IKDC score increased preoperatively from 36 to 64, the overall KOOS from 43 to 64, the NSARS score from 30 to 67, and the TAS score from 2 to 3.7. The mean MOCART score was 73; the MOCART 2.0, 69; and the 3D-MOCART, 69 and 70. The authors observed ILBO in 77% and BMELSs in 74% of patients after 60 to 120 months. Previous cartilage surgeries and osteochondral defect buildup showed higher rates of these abnormalities. Early lesions of the subchondral lamina did not predict ILBO in long-term follow-up, but BMELSs predicted later appearance with decreasing size. CONCLUSION: Subchondral changes frequently appeared in long-term MRI evaluation of patients after ACI. BMELSs showed a decreasing diameter over the years, while the size of ILBO increased in the later follow-ups. These findings did not affect the clinical outcome in the study population. However, osteoarthritis is likely to progress. The degenerative effect and influence on longer-term outcomes needs to be clarified in future studies.

Practical considerations for determination of scapular internal rotation and its relevance in reverse total shoulder arthroplasty planning.

BACKGROUND: Scapulothoracic orientation, especially scapular internal rotation (SIR) may influence range of motion in reverse total shoulder arthroplasty (RTSA) and is subjected to body posture. Clinical measurements of SIR rely on apical bony landmarks, which depend on changes in scapulothoracic orientation, while radiographic measurements are often limited by the restricted field of view (FOV) in CT scans. Therefore, the goal of this study was (1) to determine whether the use of CT scans with a limited FOV to measure SIR is reliable and (2) if a clinical measurement could be a valuable alternative. METHODS: This anatomical study analyzed the whole-body CT scans of 100 shoulders in 50 patients (32 male and 18 female) with a mean age of 61.2 ± 20.1 years (range 18; 91). (1) CT scans were rendered into 3D models and SIR was determined as previously described. Results were compared to measurements taken in 2D CT scans with a limited FOV. (2) Three apical bony landmarks were defined: (the angulus acromii (AA), the midpoint between the AA and the coracoid process tip (C) and the acromioclavicular (AC) joint. The scapular axis was determined connecting the trigonum scapulae with these landmarks and referenced to the glenoid center. The measurements were repeated with 0°, 10°, 20°, 30° and 40° anterior scapular tilt. RESULTS: Mean SIR was 44.8° ± 5.9° and 45.6° ± 6.6° in the 3D and 2D model, respectively (p < 0.371). Mean difference between the measurements was 0.8° ± 2.5° with a maximum of 10.5°. Midpoint AA/C showed no significant difference to the scapular axis at 0° (p = 0.203) as did the AC-joint at 10° anterior scapular tilt (p = 0.949). All other points showed a significant difference from the scapular axis at all degrees of tilt. CONCLUSION: 2D CT scans are reliable to determine SIR, even if the spine is not depicted. Clinical measurements using apical superficial scapula landmarks are a possible alternative; however, anterior tilt influenced by posture alters measured SIR.

A human, allogeneic cortical bone screw for distal interphalangeal joint (DIP) arthrodesis: a retrospective cohort study with at least 10 months follow-up.

INTRODUCTION: The prime requisites of a good digital arthrodesis are a painless and stable union in a proper position. Arthrodesis of the distal interphalangeal joint of the fingers is not without potential complications including nonunion, malunion, and deep tissue infections. The Shark Screw® is a human, cortical bone allograft for osteosynthesis and an alternative to metal or bioabsorbable devices in orthopedics and trauma surgery. The primary hypothesis is that the fusion and complication rate, using the Shark Screw®, is at least similar to those reported in the literature, using metal or bioabsorbable screws. MATERIAL AND METHODS: This retrospective cohort study analyzes the fusion and complication rate and the patient satisfaction of distal interphalangeal joint arthrodesis of 27 fingers with the human allogeneic cortical bone screw. Complications, Disabilities of Arm, Shoulder, and Hand Questionnaire (Quick-DASH) score and Michigan Hand Outcomes Questionnaire (MHQ) score, grip and pinch strength and fusion angle were investigated. RESULTS: The mean follow-up was 23 months. At 6 weeks after surgery, fusion was obtained for all fingers. There was no surgical complication that required revision surgery. An average fusion angle of 13.6° ± 10.7° was measured. VAS pain score decreased significantly from 6.9 before surgery to 0.14 after surgery. The Quick-DASH score decreased from 10.7 to 7.8. The MHQ score improved in all sub-scores. CONCLUSION: The complication rates, using the Shark Screw® for DIP joint arthrodesis, are lower compared to the results reported in the literature for other surgical techniques. Complications related to the human allograft cortical bone screw itself were not observed. The bone screw is completely remodeled into the host bone and further hardware removal is not necessary. LEVEL OF EVIDENCE: IV.

Target enrichment of long open reading frames and ultraconserved elements to link microevolution and macroevolution in non-model organisms.

Despite the increasing accessibility of high-throughput sequencing, obtaining high-quality genomic data on non-model organisms without proximate well-assembled and annotated genomes remains challenging. Here, we describe a workflow that takes advantage of distant genomic resources and ingroup transcriptomes to select and jointly enrich long open reading frames (ORFs) and ultraconserved elements (UCEs) from genomic samples for integrative studies of microevolutionary and macroevolutionary dynamics. This workflow is applied to samples of the African unionid bivalve tribe Coelaturini (Parreysiinae) at basin and continent-wide scales. Our results indicate that ORFs are efficiently captured without prior identification of intron-exon boundaries. The enrichment of UCEs was less successful, but nevertheless produced substantial data sets. Exploratory continent-wide phylogenetic analyses with ORF supercontigs (>515,000 parsimony informative sites) resulted in a fully resolved phylogeny, the backbone of which was also retrieved with UCEs (>11,000 informative sites). Variant calling on ORFs and UCEs of Coelaturini from the Malawi Basin produced ~2000 SNPs per population pair. Estimates of nucleotide diversity and population differentiation were similar for ORFs and UCEs. They were low compared to previous estimates in molluscs, but comparable to those in recently diversifying Malawi cichlids and other taxa at an early stage of speciation. Skimming off-target sequence data from the same enriched libraries of Coelaturini from the Malawi Basin, we reconstructed the maternally-inherited mitogenome, which displays the gene order inferred for the most recent common ancestor of Unionidae. Overall, our workflow and results provide exciting perspectives for integrative genomic studies of microevolutionary and macroevolutionary dynamics in non-model organisms.

Trematode infection affects shell shape and size in Bulinus tropicus.

Trematodes can increase intraspecific variation in the phenotype of their intermediate snail host. However, the extent of such phenotypic changes remains unclear. We investigated the influence of trematode infection on the shell morphology of Bulinus tropicus, a common host of medically important trematodes. We focused on a snail population from crater lake Kasenda (Uganda). We sampled a single homogeneous littoral habitat to minimize the influence of environmental variation on shell phenotype, and barcoded snails to document snail genotypic variation. Among the 257 adult snails analysed, 99 tested positive for trematode infection using rapid-diagnostic PCRs. Subsequently we used high-throughput amplicon sequencing to identify the trematode (co-)infections. For 86 out of the 99 positive samples trematode species delineation could discriminate among combinations of (co-)infection by 11 trematode species. To avoid confounding effects, we focused on four prevalent trematode species. We performed landmark-based geometric morphometrics to characterize shell phenotype and used regressions to examine whether shell size and shape were affected by trematode infection and the developmental stage of infection (as inferred from read counts). Snails infected by Petasiger sp. 5, Echinoparyphium sp. or Austrodiplostomum sp. 2 had larger shells than uninfected snails or than those infected by Plagiorchiida sp. Moreover, the shell shape of snails infected solely by Petasiger sp. 5 differed significantly from that of uninfected snails and snails infected with other trematodes, except from Austrodiplostomum sp. 2. Shape changes included a more protuberant apex, an inward-folded outer apertural lip and a more adapically positioned umbilicus. Size differences were more pronounced in snails with 'late' infections (>25 days) compared to earlier-stage infections. No phenotypic differences were found between snails infected by a single trematode species and those harbouring co-infections. Further work is required to assess the complex causal links between trematode infections and shell morphological alterations of snail hosts.

Drivers of diversification in freshwater gastropods vary over deep time.

Unravelling the drivers of species diversification through geological time is of crucial importance for our understanding of long-term evolutionary processes. Numerous studies have proposed different sets of biotic and abiotic controls of speciation and extinction rates, but typically they were inferred for a single, long geological time frame. However, whether the impact of biotic and abiotic controls on diversification changes over time is poorly understood. Here, we use a large fossil dataset, a multivariate birth-death model and a comprehensive set of biotic and abiotic predictors, including a new index to quantify tectonic complexity, to estimate the drivers of diversification for European freshwater gastropods over the past 100 Myr. The effects of these factors on origination and extinction are estimated across the entire time frame as well as within sequential time windows of 20 Myr each. Our results find support for temporal heterogeneity in the factors associated with changes in diversification rates. While the factors impacting speciation and extinction rates vary considerably over time, diversity-dependence and topography are consistently important. Our study highlights that a high level of heterogeneity in diversification rates is best captured by incorporating time-varying effects of biotic and abiotic factors.

Simultaneous genotyping of snails and infecting trematode parasites using high-throughput amplicon sequencing.

Several methodological issues currently hamper the study of entire trematode communities within populations of their intermediate snail hosts. Here we develop a new workflow using high-throughput amplicon sequencing to simultaneously genotype snail hosts and their infecting trematode parasites. We designed primers to amplify four snail and five trematode markers in a single multiplex PCR. While also applicable to other genera, we focused on medically and economically important snail genera within the superorder Hygrophila and targeted a broad taxonomic range of parasites within the class Trematoda. We tested the workflow using 417 Biomphalaria glabrata specimens experimentally infected with Schistosoma rodhaini, two strains of Schistosoma mansoni and combinations thereof. We evaluated the reliability of infection diagnostics, the robustness of the workflow, its specificity related to host and parasite identification, and the sensitivity to detect co-infections, immature infections and changes of parasite biomass during the infection process. Finally, we investigated its applicability in wild-caught snails of other genera naturally infected with a diverse range of trematodes. After stringent quality control the workflow allows the identification of snails to species level, and of trematodes to taxonomic levels ranging from family to strain. It is sensitive to detect immature infections and changes in parasite biomass described in previous experimental studies. Co-infections were successfully identified, opening the possibility to examine parasite-parasite interactions such as interspecific competition. Together, these results demonstrate that our workflow provides a powerful tool to analyse the processes shaping trematode communities within natural snail populations.

The fingernail clams (Bivalvia: Veneroida: Sphaeriidae) of Morocco: Diversity, distribution and conservation status.

BACKGROUND: In Morocco, many uncertainties surround the current diversity and distribution of the bivalve family Sphaeriidae. Such information, including taxonomy and conservation status, are vital for further studies to improve the knowledge of this family in Morocco and represents the first step towards the development of a national conservation plan for all freshwater bivalves.Between 2016 and 2019, several investigations were carried out to assess the diversity and distribution of Sphaeriidae in the different basins of Morocco, covering different types of habitat (lakes, springs, rivers and small ponds). The identification of specimens and their morpho-ecological features was based on morphological and morphometric analyses. The data on the general distribution of the collected species allowed the evaluation of their conservation status as well.The shell morphology and morphometric analyses revealed the existence of five species belonging to the genus Pisidium [P.casertanum (Poli, 1791), P. (cf.) personatum Malm, 1855, P.subtruncatum Malm, 1855, P.amnicum (O. F. Müller, 1774) and Pisidium sp.] and one species to the genus of Musculium [M.lacustre (O. F. Müller, 1774)]. Sphaeriidae were found in all Moroccan basins, except Bouregreg and Sakia El Hamra-Oued Eddahab Basins. The results showed that Sebou Basin was the species richest with the occurrence of the five species, while Loukkos and Sous-Massa Basins had the lowest-species richness with one species only. The conservation status of sphaeriids in Morocco was evaluated for the first time and resulted in P. (cf.) personatum and P.subtruncatum being proposed as Least Concern and Vulnerable, respectively, while the status of Regionally Extinct is suggested for both species P.amnicum and M.lacustre. NEW INFORMATION: First evaluation of the diversity and species richness of the family Sphaeriidae in Morocco with an assignment of an updated conservation status of the recorded species.

Clinical evaluation after matrix-associated autologous chondrocyte transplantation : a comparison of four different graft types.

AIMS: The aim of this retrospective study was to determine if there are differences in short-term clinical outcomes among four different types of matrix-associated autologous chondrocyte transplantation (MACT). METHODS: A total of 88 patients (mean age 34 years (SD 10.03), mean BMI 25 kg/m2 (SD 3.51)) with full-thickness chondral lesions of the tibiofemoral joint who underwent MACT were included in this study. Clinical examinations were performed preoperatively and 24 months after transplantation. Clinical outcomes were evaluated using the International Knee Documentation Committee (IKDC) Subjective Knee Form, the Brittberg score, the Tegner Activity Scale, and the visual analogue scale (VAS) for pain. The Kruskal-Wallis test by ranks was used to compare the clinical scores of the different transplant types. RESULTS: The mean defect size of the tibiofemoral joint compartment was 4.28 cm2 (SD 1.70). In total, 11 patients (12.6%) underwent transplantation with Chondro-Gide (matrix-associated autologous chondrocyte implantation (MACI)), 40 patients (46.0%) with Hyalograft C (HYAFF), 21 patients (24.1%) with Cartilage Regeneration System (CaReS), and 15 patients (17.2%) with NOVOCART 3D. The mean IKDC Subjective Knee Form score improved from 35.71 (SD 6.44) preoperatively to 75.26 (SD 18.36) after 24 months postoperatively in the Hyalograft group, from 35.94 (SD 10.29) to 71.57 (SD 16.31) in the Chondro-Gide (MACI) group, from 37.06 (SD 5.42) to 71.49 (SD 6.76) in the NOVOCART 3D group, and from 45.05 (SD 15.83) to 70.33 (SD 19.65) in the CaReS group. Similar improvements were observed in the VAS and Brittberg scores. CONCLUSION: Two years postoperatively, there were no significant differences in terms of outcomes. Our data demonstrated that MACT, regardless of the implants used, resulted in good clinical improvement two years after transplantation for localized tibiofemoral defects. Cite this article: Bone Joint Res 2021;10(7):370-379.

Third-Generation Autologous Chondrocyte Implantation at the Knee Joint Using the Igor Scaffold: A Case Series With 2-Year Follow-up.

BACKGROUND: For large, locally restricted cartilage defects in young patients, third-generation matrix-supported autologous chondrocyte implantation (ACI) with a variety of scaffolds has shown good mid- to long-term results. PURPOSE/HYPOTHESIS: This study aimed to monitor the clinical and radiological outcomes of patients who received ACI at the knee joint using the Igor scaffold (IGOR-Institute for Tissue and Organ Reconstruction) at 2-year follow-up. Our hypothesis was that there would be improvements in postoperative subjective scores and cartilage repair tissue quality. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: A total of 21 patients (12 male and 9 female) were available for 2-year follow-up after third-generation ACI using the Igor scaffold. All were clinically assessed using the Knee injury and Osteoarthritis Outcome Score (KOOS), Tegner Activity Scale, Brittberg score, International Knee Documentation Committee (IKDC) Subjective Knee Form, Noyes Sports Activity Rating Scale, and visual analog scale for pain. For morphological evaluation, the magnetic resonance observation of cartilage repair tissue (MOCART) and MOCART 2.0 scores were calculated using 3-T magnetic resonance imaging performed at 3, 6, 12, and 24 months postoperatively. Results were compared between baseline and 24 months postoperatively. RESULTS: After 2 years, the clinical and radiological scores showed good to excellent results in the majority of patients. On the IKDC, 10 patients were graded as excellent, 4 as good, 5 as fair, and 2 as severe; on the KOOS, 7 patients were graded as excellent, 8 as good, 4 as fair, and 2 as severe. From baseline to latest follow-up, visual analog scale pain scores decreased from 5.6 ± 3.2 (mean ± SD) to 1.5 ± 2; KOOS results increased from 51 ± 20.7 to 75.2 ± 15.4; and the Tegner score improved from 2.2 ± 1.8 to 4.3 ± 1.3. The MOCART and MOCART 2.0 scores were comparable at 2-year follow-up, with mean values of 74 ± 10 and 78 ± 13, respectively. Satisfactory filling and integration were found in 90.5%. Overall, 16 of 21 patients (76.1%) were satisfied with the surgery and would undergo the procedure again. CONCLUSION: Third-generation ACI using the Igor scaffold showed improvements in clinical and radiological results that were comparable with other scaffolds for patients with large traumatic or degenerative cartilage defects. Patients reported a decrease in pain and an increase in activity, with the majority reporting good results.

Population collapse in viviparid gastropods of the Lake Victoria ecoregion started before the Last Glacial Maximum.

Ecosystems of Lake Victoria and riparian communities have been strongly disrupted by the introduction of the invasive Nile perch and its fishing industry. Beyond this invasion and other recent anthropogenic stressors, the Lake Victoria ecoregion also underwent phases of pronounced aridity over the Late Pleistocene, lastly during the Last Glacial Maximum (LGM). The consequences of recent and historic environmental change have been canvassed for the adaptive radiation of haplochromine cichlids occupying the ecoregion, but their effect on freshwater invertebrate diversity remains largely unknown. Here, we use 15 microsatellite loci and approximate Bayesian computation to test whether viviparid gastropods experienced a population bottleneck during the LGM, as did cichlids. Clustering analyses support three viviparid gene pools in the Lake Victoria ecoregion, gathering specimens from 1) Lake Albert and the White Nile, 2) the Victoria Nile and Lake Kyoga and 3) Lake Victoria and tributaries. The last group contains the highest genetic diversity, but all groups have a considerable number of private alleles and are inferred to predate the LGM. Examinations of demographic history reveal a 190- to 500-fold population decline that started ~ 125-150 ka ago, thus substantially before the LGM bottleneck documented in haplochromine cichlids. Population collapses in viviparids are an order of magnitude more severe than declines in cichlids and have not been halted by the re-establishment of freshwater ecosystems since the LGM. Recent anthropogenic ecosystem deterioration is causing homogenization of previously diversified microhabitats, which may contribute to (local) extinction and enhanced gene flow among species within gene pools.

Evaluation of Meniscal Tissue after Meniscal Repair Using Ultrahigh Field MRI.

The study evaluates the meniscal tissue after primary meniscal suturing using 7-Tesla (T) magnetic resonance imaging with T2* mapping at 6 and 12 months after surgery to investigate the differences between repaired meniscal tissue and healthy meniscal tissue in the medial and lateral compartment. This prospective study included 11 patients (9m/2f) with a mean age of 30.6 years (standard deviation 9.0). Patients with a meniscal tear that was treated arthroscopically with meniscus suturing, using an all-inside technique, were included. All patients and seven healthy volunteers were imaged on a 7-T whole-body system. T2* mapping of the meniscus was applied on sagittal slices. Regions-of-interest were defined manually in the red and white zone of each medial and lateral meniscus to measure T2*-values. In the medial posterior and medial anterior horn similar T2*-values were measured in the red and white zone at 6- and 12-month follow-up. Compared with the control group higher T2*-values were found in the repaired medial meniscus. After 12-months T2*-values decreased to normal values in the anterior horn and remained elevated in the posterior horn. In the red zone of the lateral posterior horn a significant decrease in the T2*-values (from 8.2 milliseconds to 5.9 milliseconds) (p = 0.04), indicates successful repair; a tendency toward a decrease in the white zone between the 6 and 12 months follow-up was observed. In the red zone of the lateral anterior horn the T2*-values decreased significantly during follow-up and in the white zone of the lateral anterior horn T2*-values were comparable. In comparison to the control group higher T2*-values were measured at 6-months; however, the T2*-values showed comparable values in the repaired lateral meniscus after 12 months. The T2* mapping results of the current study indicated a better healing response of the red zone of the lateral posterior horn compared with the medial posterior horn.

Miocene geologic dynamics of the Australian Sahul Shelf determined the biogeographic patterns of freshwater planorbid snails (Miratestinae) in the Indo-Australian Archipelago.

The complex geological and climatic processes that have shaped the Indo-Australian Archipelago since the Cenozoic likely also gave rise to its species-rich biota. Strictly freshwater organisms might be particularly suitable for understanding the influence of these abiotic factors on their biogeography in such a insular setting as their distribution may reflect past abiotic events at large and small geographical scales. We here investigate the historical biogeography of the Miratestinae, a subfamily of Planorbidae. These freshwater gastropods are widely distributed in the eastern IAA from Australia, New Guinea, the Moluccas, and Sulawesi to the Philippines. The first comprehensive molecular phylogeny of the Miratestinae was inferred based on two mitochondrial and two nuclear genetic markers using maximum likelihood and Bayesian inference. Four species delimitation methods were applied to identify molecular operational taxonomic units (MOTUs). Divergence times were inferred using an uncorrelated lognormal relaxed-clock model by applying a taxon- and marker-specific substitution rate. Ancestral geographic ranges were estimated based on the dated phylogeny using BioGeoBEARS. The species delimitation revealed a total of 23 MOTUs, 16 of which might represent species new to science. The BioGeoBEARS analyses suggest an Australian origin for the Miratestinae at c. 22 Ma and identified jump dispersal to be the main process of colonization. The first colonization events from Australia to the IAA occurred in the Middle-Late Miocene (12-13 Ma), whereas intra-island diversification took mainly place since the Late Miocene-Pliocene. Colonization and diversification events remarkably coincide with major geologic events that shaped the geography of the region. The increasing availability of landmasses along the Sahul Shelf likely promoted stepping-stone dispersal to New Guinea, Sulawesi and the Philippines as early as the islands emerged. Major geological and climatic events such as the amalgamation of the island Sulawesi, the regional aridification in Australia or the uplift of massive mountain ranges in New Guinea likely played a considerable role for intra-island diversification.

Deep drilling reveals massive shifts in evolutionary dynamics after formation of ancient ecosystem.

The scarcity of high-resolution empirical data directly tracking diversity over time limits our understanding of speciation and extinction dynamics and the drivers of rate changes. Here, we analyze a continuous species-level fossil record of endemic diatoms from ancient Lake Ohrid, along with environmental and climate indicator time series since lake formation 1.36 million years (Ma) ago. We show that speciation and extinction rates nearly simultaneously decreased in the environmentally dynamic phase after ecosystem formation and stabilized after deep-water conditions established in Lake Ohrid. As the lake deepens, we also see a switch in the macroevolutionary trade-off, resulting in a transition from a volatile assemblage of short-lived endemic species to a stable community of long-lived species. Our results emphasize the importance of the interplay between environmental/climate change, ecosystem stability, and environmental limits to diversity for diversification processes. The study also provides a new understanding of evolutionary dynamics in long-lived ecosystems.

Multi-Omics Characterization of the 4T1 Murine Mammary Gland Tumor Model.

Background: Tumor models are critical for our understanding of cancer and the development of cancer therapeutics. The 4T1 murine mammary cancer cell line is one of the most widely used breast cancer models. Here, we present an integrated map of the genome, transcriptome, and immunome of 4T1. Results: We found Trp53 (Tp53) and Pik3g to be mutated. Other frequently mutated genes in breast cancer, including Brca1 and Brca2, are not mutated. For cancer related genes, Nav3, Cenpf, Muc5Ac, Mpp7, Gas1, MageD2, Dusp1, Ros, Polr2a, Rragd, Ros1, and Hoxa9 are mutated. Markers for cell proliferation like Top2a, Birc5, and Mki67 are highly expressed, so are markers for metastasis like Msln, Ect2, and Plk1, which are known to be overexpressed in triple-negative breast cancer (TNBC). TNBC markers are, compared to a mammary gland control sample, lower (Esr1), comparably low (Erbb2), or not expressed at all (Pgr). We also found testis cancer antigen Pbk as well as colon/gastrointestinal cancer antigens Gpa33 and Epcam to be highly expressed. Major histocompatibility complex (MHC) class I is expressed, while MHC class II is not. We identified 505 single nucleotide variations (SNVs) and 20 insertions and deletions (indels). Neoantigens derived from 22 SNVs and one deletion elicited CD8+ or CD4+ T cell responses in IFNγ-ELISpot assays. Twelve high-confidence fusion genes were observed. We did not observe significant downregulation of mismatch repair (MMR) genes or SNVs/indels impairing their function, providing evidence for 6-thioguanine resistance. Effects of the integration of the murine mammary tumor virus were observed at the genome and transcriptome level. Conclusions: 4T1 cells share substantial molecular features with human TNBC. As 4T1 is a common model for metastatic tumors, our data supports the rational design of mode-of-action studies for pre-clinical evaluation of targeted immunotherapies.