Biomedical subjects

J Xiang

Publications and source records attributed to J Xiang.

At least 19 recordsLinked to original sources

PubMed2006

HER-2/neu-gene engineered dendritic cell vaccine stimulates stronger HER-2/neu-specific immune responses compared to DNA vaccination.

HER-2/neu is a candidate for developing breast cancer-targeted immunotherapeutics. Although DNA-based and HER-2/neu transgene-modified dendritic cell (DC)-based vaccines are potent at eliciting HER-2/neu-specific antitumor immunity, there has been no side-by-side study comparing them directly. The present study utilizes an in vivo murine tumor model expressing HER-2/neu antigen to compare the efficacy between adenovirus (AdVneu)-transfected dendritic cells (DC(neu)) and plasmid DNA (pcDNAneu) vaccine. Our data showed that DC(neu) upregulated the expression of immunologically important molecules and inflammatory cytokines and partially converted regulatory T (Tr)-cell suppression through interleukin-6 (IL-6) secretion. Vaccination of DC(neu) induced stronger HER-2/neu-specific humoral and cellular immune responses than DNA vaccination, which downregulated HER-2/neu expression and lysed HER-2/neu-positive tumor cells in vitro, respectively. In two HER-2/neu-expressing tumor models, DC(neu) completely protected mice from tumor cell challenge compared to partial or no protection observed in DNA-immunized mice. In addition, DC(neu) significantly delayed breast cancer development in transgenic mice in comparison to DNA vaccine (P<0.05). Taken together, we have demonstrated that HER-2/neu-gene-modified DC vaccine is more potent than DNA vaccine in both protective and preventive animal tumor models. Therefore, DCs genetically engineered to express tumor antigens such as HER-2/neu represent a new direction in DC vaccine of breast cancer.

Adenoviridae↗

PubMed2006

Microsurgical excision of the large or giant cerebellopontine angle meningioma.

OBJECTIVE: The object of this study was to analyze the therapeutic effects of microsurgical excision in cases with the large or giant cerebellopontine angle meningioma. METHODS: We retrospectively analyzed the 56 patients who suffered from the large or giant cerebellopontine angle meningioma and underwent the microsurgical therapy, for which the suboccipital-retrosigmoidal approach was adopted in 38 cases, the temporal-occipital craniotomy, presigmoidal approach in 6 cases, the temporal-occipital craniotomy, inferotemporal tentorium cerebelli approach in 8 cases, and the temporal-occipital craniotomy, supratentorial or infratentorial allied approach in 4 cases. RESULTS: The tumors of 44 cases were all resected (Simpson I, II), with a total resection rate of 78.6%, and there was no operative mortality. After surgery, symptoms improved in 40 cases and remained unchanged in 10 cases. Among 54 cases, recrudescence was seen in 2 cases (3.7%) and being able to take care of themselves in 50 cases (92.6%) at 6 months through 6 years follow-up after surgery. CONCLUSION: A rationally selected surgical approach, a microscopic technology applied in the operation to appropriately treat and protect vein, nerve and brain stem, which can ideally excise the tumors, together can increase the survival ability of patients.

Adolescent↗

PubMed2006

Effects of RNA interference on CD80 and CD86 expression in bone marrow-derived murine dendritic cells.

To investigate whether RNA interference (RNAi) induced by small interfering RNA (siRNA) could suppress CD80 and CD86 expression in bone marrow-derived murine dendritic cells (DC). The bone marrow-derived DC of mice were separated and cultured in vitro, chemically synthesized siRNA were then transferred into the cells by LipofectAMINE 2000, and the siRNA transfection efficacy was assessed by both fluorescence microscope and flow cytometry. The mRNA expression and protein synthesis were analysed by real-time RT-PCR and flow cytometry. The cell viability of transfected DC was determined by annexin V and propidium iodine staining. Transfection of bone marrow-derived murine DC with a non-silencing FITC-labelled control siRNA demonstrated a high (71.86%) transfection efficiency without affecting cellular viability. CD80-1 siRNA was the most effective siRNA to block CD80 expression in three candidates. Similarly, CD86-3 siRNA was extraordinarily effective in repressing the expression of CD86. Cotransfection of siRNA specific to CD80 and CD86 can enhance gene silencing that is not affected by DC activation-inducing signals. CD80 and CD86 siRNA suppressed the expression of CD80 and CD86 to 31.05 +/- 2.41% and 25.43 +/- 0.85%, respectively, of the level in untreated cells (P < 0.05). siRNA is capable of triggering RNAi in bone marrow-derived DC; it can specifically and effectively knock down CD80 and CD86 gene expression. This approach is a useful tool by which costimulatory molecules of DC can be studied as well as a potential therapeutic option for allograft rejection.

Animals↗

PubMed2006

Effect of primer selection on estimates of GB virus C (GBV-C) prevalence and response to antiretroviral therapy for optimal testing for GBV-C viremia.

GB virus C (GBV-C; also called hepatitis G virus) is a common cause of infection associated with prolonged survival among HIV-infected individuals. The prevalences of GBV-C viremia vary widely in different studies, and there has been poor agreement among different laboratories performing GBV-C RNA detection in quality control studies. To determine the optimal method of measuring GBV-C RNA in clinical samples, samples obtained from 939 HIV-infected subjects were studied using reverse transcription (RT)-PCR methods amplifying four separate regions of the GBV-C genome. Primers amplifying the E2 coding region were 100% specific; however, their sensitivity was only 76.6%. In contrast, primers amplifying three additional conserved regions of the GBV-C genome (the 5' nontranslated region and the nonstructural protein-coding regions 3 and 5A) were more sensitive but produced higher rates of false-positive results. Using low-specificity primer sets influenced the significance of association between GBV-C viremia and response to antiretroviral therapy. Using a quantitative GBV-C RNA method, the GBV-C RNA concentration did not correlate with baseline or set point HIV RNA levels; however, a correlation between negative, low, and high GBV-C RNA levels and increasing reduction in HIV RNA following antiretroviral therapy was observed. Subjects with both GBV-C E2 antibody and viremia had significantly lower GBV-C RNA levels than did viremic subjects without E2 antibody. These studies demonstrate that accurate detection of GBV-C RNA by nested RT-PCR requires the use of primers representing multiple genome regions. Analyses based on testing with single primers do not lead to reliable conclusions about the association between GBV-C infection and clinical outcomes.

Anti-HIV Agents↗

PubMed2006

Epigenetic transfer of metastatic activity by uptake of highly metastatic B16 melanoma cell-released exosomes.

AIM: To investigate potential role of highly metastatic BL6-10 tumor cell-released exosomes (EXO) in transfer of metastatic activity into poorly metastatic tumor cell line F1. METHODS: The highly metastatic B16 melanoma cell line (BL6-10) was generated in our laboratory. EXO from this cell line were isolated and amount of exosomal recovered proteins was measured using Bradford assay. For phenotypic analysis BL6-10 and F1 melanoma cells were stained with FITC-conjugated anti-MHC I (H-2K(b)), MHC II (Ia(b)) and Met 72 antibodies and analyzed by flow cytometry. C57BL/6 mice (8 per group) were injected (i. v.) with 0.5 x 10(6) F1, BL6-10 and F1(EXO) melanoma cells. Lungs were removed 4 weeks after tumor cell injection, fixed in 10% neutral buffered formaldehyde and embedded in paraffin for histological analysis. RESULTS: Data revealed that BL6-10 cells expressed metastasis marker (Met 72 tumor antigen), while F1 cells did not display this cell surface marker. All mice inoculated with BL6-10 melanoma cells had numerous lung tumor colonies, while mice injected with F1 tumor cells were free of lung metastatic colonies. BL6-10 tumor cells-released EXO also expressed Met 72 tumor antigen as BL6-10 tumor cells, but in less amount. F1 tumor cells can uptake EXO from BL6-10 tumor cells and express acquired exosomal Met 72 tumor antigen. CONCLUSION: The metastatic activity of highly metastatic BL6-10 tumor cells can be transferred to poorly metastatic F1 tumor cells by uptake of highly metastatic BL6-10 tumor-released EXO.

Animals↗

PubMed2005

Combined CD4+ Th1 effect and lymphotactin transgene expression enhance CD8+ Tc1 tumor localization and therapy.

Type 1 T cells are the major components in antitumor immunity. The lack of efficient CD8(+) cytotoxic T (Tc) cell infiltration of tumors is a major obstacle to adoptive Tc-cell therapy. We have previously demonstrated that adenovirus (AdV)-mediated transgene lymphotactin (Lptn) expression by intratumoral AdVLptn injection and intravenous CD4(+) helper T (Th) cell transfer can enhance Tc-cell tumor infiltration and eradication of early stage tumors (5 mm in diameter). In this study, we generated ovalbumin (OVA)-specific Tc1 and Th1 cells in vitro by incubation of OVA-pulsed dendritic cells with naive T cells from T-cell receptor (TCR) transgenic OT I and OT II mice. We then investigated the potential synergy of Th1 help effect and Lptn transgene expression in Tc1-cell therapy of well-established OVA-expressing EG7 solid tumors (7 mm in diameter). Our data showed that a combined adoptive T-cell therapy of Th1 (2.5 x 10(6) cells per mouse) and Tc1 (5 x 10(6) cells per mouse) resulted in regression of all eight (100%) transgene Lptn expressed EG7 tumors, which is significantly higher than four from eight (50%) in AdVLptn/Tc1 group and two from eight (25%) in Tc1/Th1 group (P < 0.05). The amount of transferred Tc1 cells detected in Lptn-expressed tumors with Th1 treatment is 0.72%, which is significantly higher than those of AdVLptn (0.22%), Th1 (0.41%) and the control AdVpLpA (0.09%) treatment groups (P < 0.05). Enhanced Tc1 tumor localization may be derived from the chemotactic effect of Lptn and the proliferative effect of Th1 and Lptn. This novel therapeutic strategy with enhancement of Tc1 tumor localization in the therapy of well-established tumors may become a tool of considerable conceptual interest in the implementation of future clinical objectives.

Adenoviridae↗

PubMed2004

Volumetric estimation of functional brain regions in small children using spatially filtered magnetoencephalography: differentiating thumb from middle finger.

It is difficult to localize the somatosensory cortex in children under 6 years using magnetoencephalography (MEG) with conventional median nerve stimulation. One main reason is that MEG data recorded from small children have poor signal-to-noise ratio due to their small heads and short hands. To find a better approach, this study investigated neuromagnetic activities following finger stimulation using spatially filtered MEG. Four healthy children have been studied with a whole cortex MEG system. Electric stimulation was applied to the thumb and the middle fingers with two Digital Rings. Two trials were recorded for each hand. Focal increases of spectral power were localized using spectrogram and synthetic aperture magnetometry (SAM). A clear response at a latency of 21 ms was identified in 3 children (3/4). Dipole modeling localized the somatosensory cortex in 2 children (2/4). SAM successfully localized the somatosensory cortex in 4 children (4/4). Interestingly, the functional region estimated for the thumb was significantly larger than that of the middle finger (p < 0.01). However, it is possible that this effect is due to a larger source amplitude rather than larger source volume. Our results demonstrated that the developed approach could map the somatosensory cortex in children ranging in age from 3 to 6. To our knowledge, this is the first report using non-invasive methods to provide quantitative data indicating that the functional area of the thumb is larger than that of the middle finger in small children.

Child↗

PubMed2004

Spatiotemporal analysis of neuromagnetic activation associated with mirror reading.

Our previous report has confirmed that visually presented words can elicit neuromagnetic activities from the visual cortex, angular gyrus and Broca's area. It is not clear how the words are visuospatially transformed and recognized by the human brain. Mirror reading is characterized by reading which runs in the opposite direction to normal reading, with reversals of letters. It would be very interesting to find out if there are any neuromagnetic differences between the mirror reading and normal reading. Four right-handed healthy adults have been studied with a whole cortex Magnetoencephalography (MEG) system. The stimuli consisted of eight normal oriented words and eight inverted exclamation mark section "reversed words" (mirror-image of the words). All stimuli were randomly presented on the screen in front of the subjects using DirectX. MEG data were analyzed using both single dipole modeling and synthetic aperture magnetometry (SAM). Four responses to the reversed words were identified in all four subjects. In comparison to the normally oriented words, the reversed words elicited a stronger response at a latency of 248+/-6 ms. SAM results indicated that the reversed words invoked strong activations in the left and right parietal cortices but the normally oriented words did not. The mirror reading elicited one magnetic response which is different from that of the normal reading. The difference between the mirror reading and the normal reading in terms of neuromagnetic activation may reflect the development of novel representations for reversed words.

Adult↗

PubMed2004

Dynamic neuromagnetic responses to auditory motion: a novel index for evaluation of attention.

Our previous studies have demonstrated that there is a motion-related magnetic response evoked by moving sound but not stationary sound. The motion-related response reflects the dynamic activities of brain processing of auditory motion. This study focuses on the effect of attention on the response. Six normal hearing subjects were studied using a 151-channel whole-cortex magnetoencephalography (MEG) system. Four sound stimuli were designed: two sounds moved from left to right (rightward) and the other two sounds moved from right to left (leftward). Two sounds moving in the same direction were separated with an attention task. The attention task was a mathematical calculation. The results showed that when subjects listened to the moving sound, there was a clearly auditory motion related response at a latency of 649 +/- 14.2 ms. However, when subjects were doing the calculation while listening to the moving sounds, the motion related response completely disappeared. Strikingly, there was a dent at a latency of 600 ms. To our knowledge, this is probably the first study showing a neuromagnetic response which can be clearly elicited or inhibited by an attention task. We consider that the motion related response has the potential to be an objective index for the study of attention.

Acoustic Stimulation↗

PubMed2004

Automatic localization of epileptic zones using magnetoencephalography.

Conventional visual identification of epileptic spike is a challenging problem in the clinical application of magnetoencephalography (MEG). More importantly, the conventional method has problems of detecting other abnormalities such as high frequency oscillation in the human epileptic brain. The objective of this study was to develop a new approach using magnetic spectral analysis and spatial filtering. Twelve patients with seizure have been studied with a whole cortex MEG system. Fifteen epochs were recorded for each patient; each epoch was 120 seconds. Neuromagnetic spectrum was analyzed using a new method called accumulated spectrogram. Focal increases of spectral power were localized using synthetic aperture magnetometry (SAM). The MEG results were then compared with clinical findings. Focal increases of spectral power have been identified in all patients (12/12, 100%). The locations of the focal increases of spectral power were in agreement with dipole locations of spikes in 9 patients (9/12, 75%). A comparison between MEG results and clinical findings indicated that SAM revealed focal epileptic activities in two patients when dipole fitting failed. The results suggest that epileptic regions could be quantitatively identified and accurately localized using accumulated spectrogram and SAM. In comparison to visual identification of spike, the new approach is objective and sensitive, and provides the possibility of analyzing much wider frequency bands.

Adolescent↗

PubMed2004

Topiramate vs placebo in painful diabetic neuropathy: analgesic and metabolic effects.

BACKGROUND: Using identical methods, three simultaneous placebo-controlled trials of topiramate for painful diabetic neuropathy (PDN) did not reach significance. This independent yet concurrent placebo-controlled trial used different methods to assess topiramate efficacy and tolerability in PDN. METHODS: This 12-week, multicenter, randomized, double-blind trial included 323 subjects with PDN and pain visual analog (PVA) score of at least 40 on a scale from 0 (no pain) to 100 (worst possible pain). Topiramate (n = 214) or placebo (n = 109) was titrated to 400 mg daily or maximum tolerated dose. Short-acting rescue analgesics were permitted only during the first 6 weeks. RESULTS: Baseline characteristics were comparable between groups except for mean body weight (topiramate, 101.4 kg; placebo, 95.7 kg; p = 0.028). Twelve weeks of topiramate treatment reduced PVA scale score (from 68.0 to 46.2 mm) more effectively than placebo (from 69.1 to 54.0 mm; p = 0.038). Fifty percent of topiramate-treated subjects and 34% of placebo-treated subjects responded to treatment, defined as >30% reduction in PVA scale score (p = 0.004). Topiramate monotherapy also reduced worst pain intensity (p = 0.003 vs placebo) and sleep disruption (p = 0.020 vs placebo). Diarrhea, loss of appetite, and somnolence were the most commonly reported adverse events in the topiramate group. Topiramate reduced body weight (-2.6 vs +0.2 kg for placebo; p < 0.001) without disrupting glycemic control. CONCLUSIONS: Topiramate monotherapy reduced pain and body weight more effectively than placebo in patients with painful diabetic neuropathy.

Adolescent↗

PubMed2004

Library screening studies to investigate substrate specificity in the reaction catalyzed by cholesterol oxidase.

We tested whether it is possible to alter the substrate specificity of cholesterol oxidase for similarly sized sterols, i.e. cholesterol, beta-sitosterol and stigmasterol. Using existing X-ray crystal structures, we made a model of the predicted Michaelis complex of cholesterol and cholesterol oxidase. Based on this model, we identified five residues that are in direct contact with the steroid tail, Met58, Leu82, Val85, Met365 and Phe433. We prepared seven mutant libraries that contained the codon NYS (N = A, C, G, T; Y = C, Y; S = C, G) at one, two or three of the targeted positions by cassette mutagenesis. The libraries were screened for catalytic activity against three different sterols under k(cat)(*)/K(m)(*) conditions with 25 mol% sterol/DOPC unilamellar vesicles. The results of our screens suggest that specific packing interactions are not realized in the transition state of binding and that loss of active site water may be the predominant source of binding energy.

Base Sequence↗

PubMed2004

A density-functional study of Al-doped Ti clusters: TinAl (n = 1-13).

Equilibrium geometries, stabilities, and electronic properties of TinAl (n = 1-13) clusters have been studied by using density-functional theory with local spin density approximation and generalized gradient approximation. The ground-state structures of TinAl clusters have been obtained. The resulting geometries show that the aluminum atom remains on the surface of clusters for n < 9, but is slowly getting trapped beyond n = 9, meanwhile, the Al atom exhibits a valent transition from monovalent to trivalent. The geometric effects and electronic effects clearly demonstrate the Ti4Al cluster to be endowed with special stability. The studies on the bonds indicate the change from ionic to metalliclike.

Journal Article↗

PubMed2003

Transport of 5-aminolevulinic acid between blood and brain.

Little is known about the movement of 5-aminolevulinic acid (delta-aminolevulinic acid; ALA) between blood and brain. This is despite the fact that increases in brain ALA may be involved in generating the neuropsychiatric symptoms in porphyrias and that systemic administration of ALA is currently being used to delineate the borders of malignant gliomas. The current study examines the mechanisms involved in the movement of [(14)C]ALA across the blood-brain and blood-CSF barriers in the rat. In the adult rat, the influx rate constant (K(i)) for [(14)C]ALA movement into brain was low ( approximately 0.2 microl/g per min), was unaffected by increasing plasma concentrations of non-radioactive ALA or probenecid (an organic anion transport inhibitor) and, therefore, appears to be a diffusional process. The K(i) for [(14)C]ALA was 3-fold less than that for [(14)C]mannitol, a molecule of similar size. This difference appears to result from a lower lipid solubility rather than saturable [(14)C]ALA transport from brain to blood. The K(i) for [(14)C]ALA for uptake into the neonatal brain was 7-fold higher than in the adult. However, again, this was unaffected by increasing plasma ALA concentrations suggesting a diffusional process. In contrast, at the blood-CSF barrier, there was evidence of carrier-mediated [(14)C]ALA transport from blood to choroid plexus and blood to CSF. Both processes were inhibited by administration of non-radioactive ALA and probenecid. However, experiments in choroid plexus epithelial cell primary cultures indicated that transport in these cells was polarized with [(14)C]ALA uptake from the apical (CSF) side being about 7-fold greater than uptake from the basolateral (blood) side. In total, these results suggest that the brain is normally fairly well protected from changes in plasma ALA concentration by the very low blood-brain barrier permeability of this compound and by a saturable efflux mechanism present at the choroid plexus.

Aminolevulinic Acid↗

PubMed2002

Effective single chain antibody (scFv) concentrations in vivo via adenoviral vector mediated expression of secretory scFv.

Single chain antibodies (scFv) represent powerful interventional agents for the achievement of targeted therapeutics. The practical utility of these agents have been limited, however, by difficulties related to production of recombinant scFv and the achievement of effective and sustained levels of scFv in situ. To circumvent these limitations, we have developed an approach to express scFv in vivo. An anti-erbB2 scFv was engineered for secretion by eukaryotic cells. The secreted scFv could bind to its target and specifically suppress cell growth of erbB2-positive cells in vitro. Adenoviral vectors expressing the cDNA for the secretory scFv likewise could induce target cells to produce an anti-tumor anti-erbB2 scFv. In vivo gene transfer via the anti-erbB2 scFv encoding adenovirus also showed anti-tumor effects. Thus, by virtue of engineering a secreted version of the anti-tumor anti-erbB-2 scFv, and in vivo expression via adenoviral vector, effective concentrations of scFv were achieved. In vivo gene transfer clearly represents a powerful means to realize effective scFv-based approaches. This method will likely have applicability for a range of disorders amenable to targeted therapeutic approaches.

Adenoviridae↗

PubMed2001

Neuromagnetic spectral distribution of implicit processing of words.

Neuromagnetic spectral distributions when silently viewing words and non-words were investigated using a 151-channel whole head MEG system. The data were analyzed with synthetic aperture magnetometry (SAM). The dominant changes of spectral power were identified in the parietal and occipital cortices in 15-30 Hz, 30-60 Hz and 60-125 Hz bands following both word and non-word stimulations. The changes in Broca's and Wernicke's areas were consistently observed in 60-125 Hz bands, mainly following word stimulation. The results indicated that the occipital and parietal cortices were most probably involved in visuospatial processing of words and non-words. The language areas, particularly Broca's and Wernickes's areas, might be involved in implicit word processing such as verbal searching and/or internal speech.

Adult↗

PubMed2001

DNA array and biological characterization of the impact of the maturation status of mouse dendritic cells on their phenotype and antitumor vaccination efficacy.

We systematically investigated the impact of the relative maturation levels of dendritic cells (DCs) on their cell surface phenotype, expression of cytokines and chemokines/chemokine receptors (by DNA array and RNase protection analyses), biological activities, and abilities to induce tumor immunity. Mature DCs expressed significantly heightened levels of their antigen-presenting machinery (e.g., CD54, CD80, CD86) and numerous cytokines and chemokines/chemokine receptors (i.e., Flt-3L, G-CSF, IL-1alpha and -1beta, IL-6, IL-12, CCL-2, -3, -4, -5, -17, and -22, MIP-2, and CCR7) and were significantly better at inducing effector T cell responses in vitro. Furthermore, mice vaccinated with tumor peptide-pulsed mature DCs better survived challenge with a weakly immunogenic tumor (8 of 8 survivors) than did mice vaccinated with less mature (3 of 8 survived) or immature (0 of 8 survivors) DCs. Nevertheless, intermediate-maturity DCs expressed substantial levels of Flt-3L, IGF-1, IL-1alpha and -1beta, IL-6, CCL-2, -3, -4, -9/10, -17, and -22, MIP-2, osteopontin, CCR-1, -2, -5, and -7, and CXCR-4. Taken together, our data clearly underscore the critical nature of employing DCs of full maturity for DC-based antitumor vaccination strategies.

Animals↗

PubMed2001

Effect of coinfection with GB virus C on survival among patients with HIV infection.

BACKGROUND: Previous studies have suggested that people with human immunodeficiency virus (HIV) infection who are coinfected with GB virus C (GBV-C, or hepatitis G virus) have delayed progression of HIV disease. GBV-C is related to hepatitis C virus but does not appear to cause liver disease. METHODS: We examined the effect of coinfection with GBV-C on the survival of patients with HIV infection. We also evaluated cultures of peripheral-blood mononuclear cells infected with both viruses to determine whether GBV-C infection alters replication in vitro. RESULTS: Of 362 HIV-infected patients, 144 (39.8 percent) had GBV-C viremia in two tests. Forty-one of the patients with GBV-C viremia (28.5 percent) died during the follow-up period, as compared with 123 of the 218 patients who tested negative for GBV-C RNA (56.4 percent; P<0.001). The mean duration of follow-up for the entire cohort was 4.1 years. In a Cox regression analysis adjusted for HIV treatment, baseline CD4+ T-cell count, age, sex, race, and mode of transmission of HIV, the mortality rate among the 218 HIV-infected patients without GBV-C coinfection was significantly higher than that among the 144 patients with GBV-C coinfection (relative risk, 3.7; 95 percent confidence interval, 2.5 to 5.4). HIV replication, as measured by the detection of p24 antigen in culture supernatants, was reproducibly inhibited in cultures of peripheral-blood mononuclear cells by GBV-C coinfection. Coinfection did not alter the surface expression of HIV cellular receptors on peripheral-blood mononuclear cells, as determined by flow cytometry. CONCLUSIONS: GBV-C infection is common in people with HIV infection and is associated with significantly improved survival.

Adult↗