The bii4africa dataset of faunal and floral population intactness estimates across Africa's major land uses.

Sub-Saharan Africa is under-represented in global biodiversity datasets, particularly regarding the impact of land use on species' population abundances. Drawing on recent advances in expert elicitation to ensure data consistency, 200 experts were convened using a modified-Delphi process to estimate 'intactness scores': the remaining proportion of an 'intact' reference population of a species group in a particular land use, on a scale from 0 (no remaining individuals) to 1 (same abundance as the reference) and, in rare cases, to 2 (populations that thrive in human-modified landscapes). The resulting bii4africa dataset contains intactness scores representing terrestrial vertebrates (tetrapods: ±5,400 amphibians, reptiles, birds, mammals) and vascular plants (±45,000 forbs, graminoids, trees, shrubs) in sub-Saharan Africa across the region's major land uses (urban, cropland, rangeland, plantation, protected, etc.) and intensities (e.g., large-scale vs smallholder cropland). This dataset was co-produced as part of the Biodiversity Intactness Index for Africa Project. Additional uses include assessing ecosystem condition; rectifying geographic/taxonomic biases in global biodiversity indicators and maps; and informing the Red List of Ecosystems.

Time for a paradigm shift? Small carnivores' sensitivity highlights the importance of monitoring mid-rank predators in future global change studies.

Research Highlight: Jachowski, D. S., Marneweck, C. J., Olfenbuttel, C., & Harris, S. N. (2024). Support for the size-mediated sensitivity hypothesis within a diverse carnivore community. Journal of Animal Ecology, https://doi.org/10.1111/1365-2656.13916. A current paradigm in ecological research suggests that top predators are suitable sentinel species to identify ecosystem dysfunctions and monitor the effects of climate change. However, the adequacy of top predators to systematically take this function may be mistakenly inferred or unintentionally conflated from the fact that these species are regarded as biodiversity indicators or keystone, umbrella and flagship species in most ecosystems. Regarding terrestrial mammalian carnivores (order Carnivora), some researchers recently suggested that the smaller species likely possess a higher sensitivity to environmental changes than large carnivores because of their biological attributes and their intermediate position in food webs. To test this hypothesis, Jachowski et al. (2024) used camera trapping followed by occupancy and structural equation modelling to explore the dynamics of a diverse carnivore community and the factors that influence them. Their results confirmed that small carnivores are more sensitive to habitat changes and are interconnected by a greater number of significant pathways compared with larger carnivores. This support for the size-mediated sensitivity hypothesis strengthens the proposition that small carnivores (and other mid-rank predators) are ideal sentinel species for monitoring the effects of the wide range of contemporary and future environmental changes. Time will tell whether this new 'middle-out ecology' paradigm will be considered in future global change studies.

Crucial choices in a global health crisis: Revealing the demand and willingness to pay for a hypothetical monkeypox vaccine - the PREVENT study.

Background: The latent monkeypox outbreak has become the most emergent public health challenge globally. This study was conducted to assess the acceptability, and willingness to take and pay for a hypothetical Monkeypox vaccine among the Vietnamese general public as well as investigate preference for individual vaccine attributes. Methods: An online cross-sectional study was conducted using snowball sampling among 842 respondents in Vietnam in 2022. A Discrete choice experiment (DCE) on preference for six major attributes of vaccine: effectiveness, immunity duration, side effects, mortality rate, restriction, and the cost was applied. Results: Fear of the impact of monkeypox on public health and the economy, vaccine service satisfaction and responsibility to the community were the most weighted factors in the decision to take a hypothetical monkeypox vaccine. Two-thirds of participants were willing to take the vaccine, while insufficient information on monkeypox and the vaccine were the main reasons for vaccine hesitancy. For vaccine attributes, the mortality rate after seven days of vaccination was the most weighted while cost was the least influential attribute. Factors associated with willingness to take and to pay for the monkeypox vaccine included knowledge of transmission, geographical location, service satisfaction, and risk of infection, while financial burden and fear of vaccine were major drivers of hesitancy. Conclusion: Our findings underline an urgent need for effective information dissemination through social media and counseling. The implementation of nationwide monkeypox vaccination requires prioritization and support for high-risk groups as well as consideration for the country's financial resources.

Efficient control of the fungal pathogens Colletotrichum gloeosporioides and Penicillium digitatum infecting citrus fruits by native soilborne Bacillus velezensis strains.

Destruction of citrus fruits by fungal pathogens during preharvest and postharvest stages can result in severe losses for the citrus industry. Antagonistic microorganisms used as biological agents to control citrus pathogens are considered alternatives to synthetic fungicides. In this study, we aimed to identify fungal pathogens causing dominant diseases on citrus fruits in a specialized citrus cultivation region of Vietnam and inspect soilborne Bacillus isolates with antifungal activity against these pathogens. Two fungal pathogens were characterized as Colletotrichum gloeosporioides and Penicillium digitatum based on morphological characteristics and ribosomal DNA internal transcribed spacer sequence analyses. Reinfection assays of orange fruits confirmed that C. gloeosporioides causes stem-end rot, and P. digitatum triggers green mold disease. By the heterologous expression of the green fluorescent protein (GFP) in C. gloeosporioides using Agrobacterium tumefaciens-mediated transformation, we could observe the fungal infection process of the citrus fruit stem-end rot caused by C. gloeosporioides for the first time. Furthermore, we isolated and selected two soilborne Bacillus strains with strong antagonistic activity for preventing the decay of citrus fruits by these pathogens. Molecular analyses of 16 S rRNA and gyrB genes showed that both isolates belong to B. velezensis. Antifungal activity assays indicated that bacterial culture suspensions could strongly inhibit C. gloeosporioides and P. digitatum, and shield orange fruits from the invasion of the pathogens. Our work provides a highly effective Bacillus-based preservative solution for combating the fungal pathogens C. gloeosporioides and P. digitatum to protect citrus fruits at the postharvest stages.

Dual roles of oxostephanine as an Aurora kinase inhibitor and angiogenesis suppressor.

The Aurora kinases, including Aurora A, B and C, play critical roles in cell division. They have been found overexpressed in a number of types of cancer and may thus be potential targets in cancer therapy. Several Aurora kinase inhibitors have been identified and developed. Some of these have been used in clinical trials and have exhibited certain efficacy in cancer treatment. However, none of these has yet been applied clinically due to the poor outcomes. Oxostephanine is an aporphine alkaloid isolated from several plants of the genus Stephania. This compound has been reported to inhibit Aurora kinase activity in kinase assays and in cancer cells. The present study aimed to investigate the real­time effects of oxostephanine extracted from Stephania dielsiana Y.C. Wu leaves on the growth of an ovarian cancer cell line (OVCAR­8, human ovarian carcinoma); these effects were compared to those of the well­known Aurora kinase inhibitor, VX­680. The effects of oxostephanine on stromal cells, as well as endothelial cells were also examined. The results demonstrated that oxostephanine was an Aurora kinase inhibitor through the prevention of histone H3 phosphorylation at serine 10, the mislocalization of Aurora B and the induction of aneuploidy. Moreover, this substance was selectively cytotoxic to human umbilical vein endothelial cells (hUVECs), whereas it was less cytotoxic to human fibroblasts and umbilical cord­derived mesenchymal stem cells. In addition, this compound significantly attenuated the migration and tube formation ability of hUVECs. Taken together, the present study demonstrates that oxostephanine plays dual roles in inhibiting Aurora kinase activity and angiogenesis. Thus, it may have potential for use as a drug in cancer treatment.

Bacterial Expression and Purification of Human Matrix Metalloproteinase-3 using Affinity Chromatography.

Matrix metalloproteinases (MMPs) belong to the family of metzincin proteases with central roles in extracellular matrix (ECM) degradation and remodeling, as well as interactions with several growth factors and cytokines. Overexpression of specific MMPs is responsible in several diseases such as cancer, neurodegenerative diseases, and cardiovascular disease. MMPs have been the center of attention recently as targets to develop therapeutics that can treat diseases correlated to MMP overexpression. To study the MMP mechanism in solution, more facile and robust recombinant protein expression and purification methods are needed for the production of active, soluble MMPs. However, the catalytic domain of most MMPs cannot be expressed in Escherichia coli (E. coli) in soluble form due to lack of posttranslational machinery, whereas mammalian expression systems are usually costly and have lower yields. MMP inclusion bodies must undergo the tedious and laborious process of extensive purification and refolding, significantly reducing the yield of MMPs in native conformation. This paper presents a protocol using Rosetta2(DE3)pLysS (hereafter referred to as R2DP) cells to produce matrix metalloproteinase-3 catalytic domain (MMP-3cd), which contains an N-terminal His-tag followed by pro-domain (Hisx6-pro-MMP-3cd) for use in affinity purification. R2DP cells enhance the expression of eukaryotic proteins through a chloramphenicol-resistant plasmid containing codons normally rare in bacterial expression systems. Compared to the traditional cell line of choice for recombinant protein expression, BL21(DE3), purification using this new strain improved the yield of purified Hisx6-pro-MMP-3cd. Upon activation and desalting, the pro domain is cleaved along with the N-terminal His-tag, providing active MMP-3cd for immediate use in countless in vitro applications. This method does not require expensive equipment or complex fusion proteins and describes rapid production of recombinant human MMPs in bacteria.

MOXAIC: A classification of major maxillofacial wounds, concerning 310 cases.

INTRODUCTION: Although classification for facial fractures have been extensively described in the literature, corresponding systems for major maxillofacial wounds (MMW) are few. We would like to present MOXAIC: a new classification system for MMW. MATERIAL AND METHODS: A retrospective study of 310 patients with MMW who underwent emergency operation between January 2005 and December 2016. MMW was defined as a facial wound longer than 10 cm, which includes damage to the craniofacial bone or other important facial structures such as the carotid arteries, facial nerves, parotid gland, Stensen's duct, or the eye. All the patients were followed at least 36 months. RESULT: Based on the shape of the wound, the severity, and the mechanism of injury we were able to classify the MMW into five types: O, X, A, I, C. For each wound type we then looked at the treatment required and the outcome, objectively classified as good, satisfactory, or poor, concerning anatomical correction, aesthetics, and function. + Type OCircumferential wound: 81.6% result good. + Type X-Oblique wound: only 48.1% good, despite initial multidisciplinary approach. + Type A-Transverse facial wound: 78.1% good. + Type I-Direct wound: Immediate airway management and hemorrhage control are important. 48.8% good. + Type CCut wound: 88.1% good. The above classification was named MOXAIC which is a mnemonic of 'Maxillofacial' and the five wound types: O, X, A, I, C. CONCLUSION: This classification is highly reproducible, easy to use, and allows quick treatment work up and prognosis. However, this classification requires further specialist review and study.

Effect of pH of coating solution on the adhesion strength and corrosion resistance of hydroxyapatite and octacalcium phosphate coated WE43 alloy.

Hydroxyapatite (HAp) and octacalcium phosphate (OCP) layers were formed on Mg- 4mass% Y- 3mass% rare earth (WE43) alloy by a chemical solution deposition method at various pH values of pH 5.5, 6.2, 7.5, and 8.6. Adhesion strength of HAp and OCP layers was evaluated before and after immersing in a medium for 14 days by a pull-off test. The corrosion resistance of these coatings was measured by polarization tests performed in a simulated body fluid (SBF). XRD analysis demonstrated that HAp coating layers were formed at pH 7.5 and 8.6, while OCP coating layers were formed at pH 5.5 and 6.2. Adhesion test results showed that the as-coated pH7.5-HAp layer had the highest adhesion strength of 8.6 MPa, which was attributed to the very dense structure of the coating layer. The as-coated pH8.6-HAp layer showed the adhesion strength of 6.5 MPa. The adhesion strength of the as-coated pH5.5- and pH6.2-OCP layers was 3.9 and 7.1 MPa, respectively, that was governed by the thick and fragile property of the layers. After immersing in the medium for 14 days, the adhesion strength of pH7.5- and pH8.6-specimens decreased to 5.8 and 5.6 MPa, respectively. The pitting corrosion and formation of Mg(OH)2 under the HAp layers were responsible for the decrease of adhesion strength. The polarization tests in SBF at 37 °C showed that the corrosion current density decreased with the HAp and OCP coatings, indicating the improvement of the corrosion resistance of WE43 alloy. The HAp coatings improved the corrosion resistance more efficiently than the OCP coatings.

Complete Vision Loss after Laparoscopic Hysterectomy.

Postoperative vision loss (POVL) is a rare but devastating complication that has only recently been reported following laparoscopic surgery. We present the case of a 34-year-old gravida 6 para 4 female who experienced POVL following an uncomplicated laparoscopic hysterectomy. Operating time was 174 minutes, and EBL was 75 mL. After surgery, she complained of complete vision loss with no light perception. No cerebral hemorrhage or ischemia was detected on imaging. Funduscopic exam revealed no structural abnormalities. On postoperative day 7, she received an IV methylprednisolone taper. The following morning, she reported mild light perception. Later that night, she reported a partial return of visual acuity and was discharged home. At her 2-week postoperative visit, her vision had returned to baseline. POVL is an emergency and prompt evaluation should be initiated to optimize outcome.

A Structured, Hybrid Cystoscopy Simulation Curriculum for Obstetrics/Gynecology Residents.

OBJECTIVES: This study aimed to assess the subjective and objective cystoscopy skills of gynecology residents before and after implementation of a comprehensive simulation curriculum. METHODS: Residents in an obstetrics/gynecology program at a single academic institution participated in a 6-week cystoscopy simulation course. Residents attended an initial orientation and didactic presentation, and then weekly 20- to 30-minute training sessions using a water balloon model and a virtual reality simulator. Pretesting and posttesting were performed, including (1) subjective self-assessments, (2) a written quiz, and (3) objective structured assessment of technical skills. Paired t test was used to assess the differences in residents' objective assessment scores before and after simulation training. RESULTS: A total of 16 residents were recruited. Residents reported significant subjective improvements in comfort, confidence, and proficiency in performing cystoscopy after completing the curriculum. Their overall objective assessment improved, including written quiz scores (mean score, 49% vs 78%; P < 0.001), efficiency in cystoscope assembly (means score, 0 vs 4; P < 0.001), overall bladder survey score (56% vs 86%, P < 0.001), and global rating score (mean score, 15 vs 24; P < 0.001). Residents tended to prefer the water-balloon model compared with the virtual reality simulator (75% vs 25%). CONCLUSIONS: Obstetrics/gynecology residents demonstrated significant improvements in subjective and objective measures of skill after completion of a 6-week cystoscopy simulation training course using a combination of low- and high-fidelity models. Although universal cystoscopy at the time of hysterectomy for benign indications remains controversial, cystoscopy simulation should remain an important part of resident training.

Metalloproteinases and Their Inhibitors: Potential for the Development of New Therapeutics.

The metalloproteinase (MP) family of zinc-dependent proteases, including matrix metalloproteinases (MMPs), a disintegrin and metalloproteases (ADAMs), and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs) plays a crucial role in the extracellular matrix (ECM) remodeling and degradation activities. A wide range of substrates of the MP family includes ECM components, chemokines, cell receptors, and growth factors. Metalloproteinases activities are tightly regulated by proteolytic activation and inhibition via their natural inhibitors, tissue inhibitors of metalloproteinases (TIMPs), and the imbalance of the activation and inhibition is responsible in progression or inhibition of several diseases, e.g., cancer, neurological disorders, and cardiovascular diseases. We provide an overview of the structure, function, and the multifaceted role of MMPs, ADAMs, and TIMPs in several diseases via their cellular functions such as proteolysis of other cell signaling factors, degradation and remodeling of the ECM, and other essential protease-independent interactions in the ECM. The significance of MP inhibitors targeting specific MMP or ADAMs with high selectivity is also discussed. Recent advances and techniques used in developing novel MP inhibitors and MP responsive drug delivery tools are also reviewed.

Patterns of nest box sharing in woodland dormice (Graphiurus murinus): Evidence for intra-sexual tolerance and communal nesting.

Nest sharing in rodents and other vertebrates is believed to be promoted by the gain of thermoregulatory advantages, reproductive imperatives or to be a by-product of other benefits to group living. Here, we studied the patterns of nest box sharing over a yearly cycle in a wild population of the arboreal, nocturnal and heterothermic woodland dormouse (Graphiurus murinus). We evaluated whether the timing of dormouse aggregations and group composition were compatible with thermoregulatory or reproductive imperatives. Nest box sharing between two or more dormice was observed throughout the year and was not correlated with monthly minimum temperatures. Therefore, the observed aggregations could not be exclusively related to the gain of a thermoregulatory advantage. All-male and all-female aggregations were more frequent during the mating season, following which some females seemed to engage in communal nesting and continued to share nests with some of their offspring well beyond the weaning period. Our data suggest that nest sharing was only partly affected by basic reproductive needs. We hypothesize that dormouse aggregations were ultimately favoured by a high level of intra-sexual tolerance elicited by the presence of numerous kin-related and familiar individuals in the population. This seems to stem from the high inter-individual spatial overlaps and the promiscuous mating system previously reported in this species. Further studies focusing on natural resting sites and genetic relatedness between individuals are needed to confirm our findings and test our hypothesis.

Competition and specialization in an African forest carnivore community.

Globally, human activities have led to the impoverishment of species assemblages and the disruption of ecosystem function. Determining whether this poses a threat to future ecosystem stability necessitates a thorough understanding of mechanisms underpinning community assembly and niche selection. Here, we tested for niche segregation within an African small carnivore community in Kibale National Park, Uganda. We used occupancy modeling based on systematic camera trap surveys and fine-scale habitat measures, to identify opposing preferences between closely related species (cats, genets, and mongooses). We modeled diel activity patterns using kernel density functions and calculated the overlap of activity periods between related species. We also used co-occupancy modeling and activity overlap analyses to test whether African golden cats Caracal aurata influenced the smaller carnivores along the spatial and/or temporal axes. There was some evidence that related species segregated habitat and activity patterns. Specialization was particularly strong among forest species. The cats and genets partitioned habitat, while the mongooses partitioned both habitat and activity period. We found little evidence for interference competition between African golden cats and other small carnivores, although weak interference competition was suggested by lower detection probabilities of some species at stations where African golden cats were present. This suggests that community assembly and coexistence in this ecosystem are primarily driven by more complex processes. The studied carnivore community contains several forest specialists, which are typically more prone to localized extinction. Preserving the observed community assemblage will therefore require the maintenance of a large variety of habitats, with a particular focus on those required by the more specialized carnivores.

Targeted delivery of nano-PTX to the brain tumor-associated macrophages.

Nanoparticles containing mixed lipid monolayer shell, biodegradable polymer core and rabies virus glycoprotein (RVG) peptide as brain targeting ligand, were developed for brain targeted delivery of paclitaxel (PTX) to treat malignant glioma. RVG conjugated PTX loaded NPs (RVG-PTX-NPs) had the desirable size (~140 nm), narrow size distribution and spherical shape. RVG-PTX-NPs showed poor uptake by neurons and selective targeting to the brain tumor associated macrophages (TAMs) with controlled release and tumor specific toxicity. In vivo studies revealed that RVG-PTX-NPs were significant to cross the blood-brain barrier (BBB) and had specific targeting to the brain. Most importantly, RVG-PTX-NPs showed effectiveness for anti-glioma therapy on human glioma of mice model. We concluded that RVG-PTX-NPs provided an effective approach for brain-TAMs targeted delivery for the treatment of glioma.

Revisiting the phylogeography and demography of European badgers (Meles meles) based on broad sampling, multiple markers and simulations.

Although the phylogeography of European mammals has been extensively investigated since the 1990s, many studies were limited in terms of sampling distribution, the number of molecular markers used and the analytical techniques employed, frequently leading to incomplete postglacial recolonisation scenarios. The broad-scale genetic structure of the European badger (Meles meles) is of interest as it may result from historic restriction to glacial refugia and/or recent anthropogenic impact. However, previous studies were based mostly on samples from western Europe, making it difficult to draw robust conclusions about the location of refugia, patterns of postglacial expansion and recent demography. In the present study, continent-wide sampling and analyses with multiple markers provided evidence for two glacial refugia (Iberia and southeast Europe) that contributed to the genetic variation observed in badgers in Europe today. Approximate Bayesian computation provided support for a colonisation of Scandinavia from both Iberian and southeastern refugia. In the whole of Europe, we observed a decline in genetic diversity with increasing latitude, suggesting that the reduced diversity in the peripheral populations resulted from a postglacial expansion processes. Although MSVAR v.1.3 also provided evidence for recent genetic bottlenecks in some of these peripheral populations, the simulations performed to estimate the method's power to correctly infer the past demography of our empirical populations suggested that the timing and severity of bottlenecks could not be established with certainty. We urge caution against trying to relate demographic declines inferred using MSVAR with particular historic or climatological events.

Using genetic methods to investigate dispersal in two badger (Meles meles) populations with different ecological characteristics.

Understanding the dispersal behaviour of a species is important for understanding its ecology and evolution. Dispersal in the Eurasian badger (Meles meles) is believed to be very limited, with social groups forming primarily through the retention of offspring. However, most of our knowledge of dispersal in this species comes from studies of high-density populations in the United Kingdom, where badgers are atypical in their behaviour, physiology, ecology and prey specialization. In this study we use genetic methods to compare dispersal patterns in a British and a Swiss population that differ in their ecology and demography. We present well-supported evidence that badgers disperse much further in the low-density continental population, where dispersal may also be female biased. Limited dispersal thus seems not to be an intrinsic behavioural characteristic of the species. Rather, dispersal patterns seem to vary depending on population demography and, ultimately, habitat quality and characteristics. This could have important management consequences, as dispersal can affect the impact of local extinction, and host dispersal has a particularly important role in disease transmission. Even though concentrated studies of a species in a single location may not provide representative data for the species, there are few mammalian studies that compare demography and dispersal patterns across contrasting habitats. Our results provide an example of phenotypic plasticity and suggest that dispersal is determined by the interaction of individual, social and environmental factors that may differ between populations.

Common structure of rare replication-deficient E1-positive particles in adenoviral vector batches.

The use of the PER.C6 adenovirus packaging cell line in combination with a designated vector plasmid system, whereby the cell line and vector with E1 deleted have no sequence overlap, eliminates the generation of replication-competent adenovirus during vector production. However, we have found cytopathic effect (CPE)-inducing particles in 2 out of more than 40 large-scale manufacturing lots produced in PER.C6 cells. The CPE inducer was detected at a frequency of 1 event in 7.5 x 10(12) vector particles. Despite amplification, it was not readily purified, indicating that the agent itself is replication deficient and requires the parental recombinant adenovirus serotype 5 (rAd5) vector for replication and packaging. Therefore, we designated the agent as a helper-dependent E1-positive region containing viral particle (HDEP). Here, we report the molecular structure of the HDEP genome, revealing an Ad comprised of E1 sequences derived from PER.C6 cells flanked by inverted terminal repeat, packaging signal, and transgene sequences. These sequences form a palindromic structure devoid of E2, E3, E4, and late genes. Since only 5 bp were shared between E1 sequences in the PER.C6 genome and viral vector sequences, the data strongly suggested that insertion of genomic DNA into an adenoviral genome had occurred essentially via nonhomologous recombination. HDEPs have been found in unrelated virus batches and appear to share a common structure that may explain their mechanism of generation. This finding allowed development of an HDEP assay to screen batches of rAd5 produced on the PER.C6 cell line and resulted in detection of seven HDEP agents from four different transgene-virus vector constructs in separate batches of Ad.

Clinical effect of “ Hypertension treating tea”

In the national Institute of traditional medicine, 60 subjects were diagnosed as hypertension of mild and moderate degree. The highest rate 34/60 (56,7%) of patients is at the group of 45-49 years old of age and 16/60 patients (22,6%) at 75-90. Among them there are 55% female (33/60) and 45% male (27/60). Results shows that in Hypertension treating tea, there is no toxic effect, the decrease of blood pressure manifested dramatically at the mild and moderate stage of hypertension, especially for systolic pressure; in addition, some biochemical indication of blood and proteinuria were improved positively, there is no side effect. The hypertension treating tea is used simply and easily, with its components of domestic materia medica, it is nonexpensive and propriate in primary care

A single short stretch of homology between adenoviral vector and packaging cell line can give rise to cytopathic effect-inducing, helper-dependent E1-positive particles.

An undesirable byproduct from recombinant adenoviral vectors is the emergence of replication competent adenovirus (RCA) that result from rare homologous recombination events between the viral E1-containing (permissive) mammalian host cell genome and the virus itself, restoring the E1 gene to the viral genome. To reduce or eliminate the problem of RCA, we evaluated production of a first generation Ad5 vector (Ad5FGF4) in the cell line PER.C6. This E1-transformed human cell line contains only Ad5 nucleotides 459-3510, which precludes double crossover-type homologous recombination because the Ad5FGF-4 only contains 5' Ad5 sequence up to nucleotide 453. The Ad5FGF4 vector does, however, retain 177 nucleotides of the 3' end of the E1B-55K gene that are also present in PER.C6. With only this single region of homology between vector and cell line, we were surprised to detect virus-specific cytopathic effects (CPE) in our cell-based assay for RCA. This CPE-inducing agent was amplified in nonpermissive A549 cells but also supported amplification of the parental Ad5FGF-4. Because we were unable to isolate the CPE-inducing agent in pure form we first identified it as atypical RCA. Polymerase chain reaction (PCR) and Southern blot experiments identified viral DNA segments in which recombination had occurred between the 177 nucleotides of E1B present in both Ad5FGF-4 and PER.C6. The atypical RCA genomes contain a copy of the original (PGK promoter-E1 gene carrying) plasmid used in the construction of the PER.C6 cell line and they retained the parental FGF-4 transgene. However, significant deletions occurred within the recombined genomes in compensation for the large insertion from PER.C6 sequences and resulted in the loss of essential viral genes. This deletion renders these recombinant viruses replication defective, requiring helper functions from remaining parental Ad5FGF-4 for amplification. These atypical RCA entities may be more properly designated as helper-dependent E1-positive particles (HDEPs). This finding shows the importance of avoiding the use of "nonmatched" vectors where any overlap exists between the recombinant vector and E1 sequences in the packaging cell line. The cloning of the FGF-4 transgene into an adenoviral vector specifically "matched" for PER.C6 (lacking the 177 nucleotide region of homology) has allowed extensive virus propagation (Ad5.1FGF-4) with no CPE- or HDEP-like events yet detected.