Step into the era of large multimodal models: a pilot study on ChatGPT-4V(ision)'s ability to interpret radiological images.

BACKGROUND: The introduction of ChatGPT-4V's 'Chat with images' feature represents the beginning of the era of large multimodal models (LMMs), which allows ChatGPT to process and answer questions based on uploaded images. This advancement has the potential to transform how surgical teams utilize radiographic data, as radiological interpretation is crucial for surgical planning and postoperative care. However, a comprehensive evaluation of ChatGPT-4V's capabilities in interpret radiological images and formulating treatment plans remains to be explored. PATIENTS AND METHODS: Three types of questions were collected: (1) 87 USMLE-style questions, submitting only the question stems and images without providing options to assess ChatGPT's diagnostic capability. For questions involving treatment plan formulations, a five-point Likert scale was used to assess ChatGPT's proposed treatment plan. The 87 questions were then adapted by removing detailed patient history to assess its contribution to diagnosis. The diagnostic performance of ChatGPT-4V was also tested when only medical history was provided. (2) We randomly selected 100 chest radiography from the ChestX-ray8 database to test the ability of ChatGPT-4V to identify abnormal chest radiography. (3) Cases from the 'Diagnose Please' section in the Radiology journal were collected to evaluate the performance of ChatGPT-4V in diagnosing complex cases. Three responses were collected for each question. RESULTS: ChatGPT-4V achieved a diagnostic accuracy of 77.01% for USMLE-style questions. The average score of ChatGPT-4V's treatment plans was 3.97 (Interquartile Range: 3.33-4.67). Removing detailed patient history dropped the diagnostic accuracy to 19.54% (P<0.0001). ChatGPT-4V achieved an AUC of 0.768 (95% CI: 0.684-0.851) in detecting abnormalities in chest radiography, but could not specify the exact disease due to the lack of detailed patient history. For cases from 'Diagnose Please' ChatGPT provided diagnoses consistent with or very similar to the reference answers. CONCLUSION: ChatGPT-4V demonstrated an impressive ability to combine patient history with radiological images to make diagnoses and directly design treatment plans based on images, suggesting its potential for future application in clinical practice.

Fucoidan reduced the invasion of oral squamous cell carcinoma cells and modified their effects to macrophages.

Fucoidan is a complex of polysaccharides showing antitumor and immunomodulation properties. Our previous studies found its regulation to myeloid immune cells, including macrophages. Aberrant infiltration and functions of macrophages are commonly found in oral squamous cell carcinoma (OSCC). In this study, we analyzed the effects of fucoidan on invasion of OSCC cells, and their regulation to macrophages, trying to evaluate its role as a potential therapy for OSCC. CAL27 and THP-1-derived macrophages were used as models for OSCC cells and tumor-infiltrated macrophages in the in vitro study, respectively. The effects of fucoidan on invasion of OSCC cells and their recruitment to macrophages were analyzed by transwell assay. KIF4A siRNA transfection was performed to investigate its role in fucoidan-modulated OSCC cells invasion. CCL3-neutralizing antibody was added into the conditioned medium of OSCC cells to evaluate its role in fucoidan-mediated macrophages recruitment and re-education. Fucoidan reduced the invasive potential of CAL27 cells with a decrease of MMP-2 and KIF4A transcription. KIF4A knockdown in CAL27 cells led to decreased invasion and MMP-2 expression. The conditioned medium of fucoidan-treated CAL27 cells promoted recruitment and inflammatory cytokines secretion on THP-1-derived macrophages. Further analysis found that fucoidan increased CCL3 production in CAL27 cells. Blocking CCL3 expression reversed the effects of fucoidan on macrophage recruitment and re-education. Our study found that fucoidan regulated the invasion of OSCC cells and also their recruiting and re-educating effects on macrophages, suggesting it could be a complementary approach in the treatment of OSCC.

Phylogeny and biogeography of Poecilia (Cyprinodontiformes: Poeciliinae) across Central and South America based on mitochondrial and nuclear DNA markers.

Poeciliids are a diverse group of small Neotropical fishes, and despite considerable research attention as models in ecology and evolutionary biology, our understanding of their biogeographic and phylogenetic relationships is still limited. We investigated the phylogenetic relationships of South and Central American Poecilia, by examining 2395 base pairs of mitochondrial DNA (ATPase 8/6, COI) and nuclear DNA (S7) for 18 species across six subgenera. Fifty-eight novel sequences were acquired from newly collected specimens and 20 sequences were obtained from previously published material. Analyses of concatenated and partitioned mitochondrial DNA and nuclear DNA sets resulted in a well-supported phylogeny that resolved several monophyletic groups corresponding to previously hypothesized subgenera and species complexes. A divergence-dating analysis supported the hypothesis of the genus Poecilia dispersing into Central America in the early Pliocene (ancestors of Psychropoecilia+Allopoecilia+Mollienesia: 7.3-2.0Mya) from predominantly South America. Subsequently, one lineage (subgenus Allopoecilia: 5.1-1.3Mya) expanded deeper into South America from Lower-Central America, and one lineage expanded from Nuclear-Central America into South America (subgenus Mollienesia: 0.71-0.14Mya). The subgenus Mollienesia diverged into three monophyletic groups that can be identified by nuptial male dorsal fin morphology and inner jaw dentition. A subclade of the unicuspid short-fins (subgenus Mollienesia) was the lineage that expanded into South America during the middle Pleistocene. Species in this subclade are now distributed across northern South America, where they are partially sympatric with Allopoecilia. However the P. (A.) caucana complex was not monophyletic, with P. (A.) wandae clustering in the Mollienesia subclade that expanded into South America. It is apparent that characters (body size, scale count, pigmentation, and gonopodium morphology) used to define the P. (A.) caucana complex are homoplastic and suggestive of rapid convergence in northern South America. Our improved taxon sampling and divergence-time calibration allowed for insights into the timing and direction of dispersals, and provides an improved understanding of the biogeographic history of an enigmatic group of fishes. Furthermore, we provided strong evidence for the monophyly of the subgenus Mollienesia and further substantiated its species complexes; therefore, we advise a taxonomic re-evaluation for the P. (A.) caucana complex to maintain monophyly of both Mollienesia and Allopoecilia.

Complete mitochondrial genome of Florida pompano Trachinotus carolinus (Teleostei, Carangidae).

The complete mitochondrial genome sequence of Florida pompano Trachinotus carolinus was determined by the overlapped polymerase chain reaction. The complete mitochondrial DNA sequence is 16,544 bp in length. It consists of 13 protein-coding genes, 22 transfer RNA genes, two rRNA genes and two non-coding regions. Overall base composition of its mitochondrial genome is estimated to be 28.68% for A, 16.27% for G, 26.00% for T, 29.06% for C, respectively, with a high A+T content (54.68%). The control region contains three conserved sequence blocks, a termination-associated sequence and a TATA box. The sequence data of T. carolinus can provide useful information for the studies on population structure, molecular systematic, stock evaluation and conservation genetics. It is also helpful to develop the rational management strategies for T. carolinus resource.

Differential protein expression of Caco-2 cells treated with selenium nanoparticles compared with sodium selenite and selenomethionine.

The study was designed to determine the differential protein expression of Caco-2 cells treated with different forms of selenium including sodium selenite, selenomethionine (Se-Met), and selenium nanoparticles (nano-Se). Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS) were used to identify the differentially expressed proteins. The results indicated that seven protein spots, ubiquitin-conjugating enzyme E2 (E2), glutathione synthetases (GS), triosephosphate isomerase (TSP), T-complex protein 1 subunit zeta (TCPZ), lamin-B1, heterogeneous nuclear ribonucleoprotein F (hnRNP F), and superoxide dismutase [Cu-Zn] (Cu, Zn-SOD) were significantly different among all the groups. According to the order of control, sodium selenite, Se-Met, and Nano-Se, the expression levels of two proteins (E2 and GS) increased and the other differential proteins were reverse. Except for E2, there were no significant differences in other protein expressions between the groups treated with nano-Se and Se-Met.

Genetic variations in two seahorse species (Hippocampus mohnikei and Hippocampus trimaculatus): evidence for middle Pleistocene population expansion.

Population genetic of seahorses is confidently influenced by their species-specific ecological requirements and life-history traits. In the present study, partial sequences of mitochondrial cytochrome b (cytb) and control region (CR) were obtained from 50 Hippocampus mohnikei and 92 H. trimaculatus from four zoogeographical zones. A total of 780 base pairs of cytb gene were sequenced to characterize mitochondrial DNA (mtDNA) diversity. The mtDNA marker revealed high haplotype diversity, low nucleotide diversity, and a lack of population structure across both populations of H. mohnikei and H. trimaculatus. A neighbour-joining (NJ) tree of cytb gene sequences showed that H. mohnikei haplotypes formed one cluster. A maximum likelihood (ML) tree of cytb gene sequences showed that H. trimaculatus belonged to one lineage. The star-like pattern median-joining network of cytb and CR markers indicated a previous demographic expansion of H. mohnikei and H. trimaculatus. The cytb and CR data sets exhibited a unimodal mismatch distribution, which may have resulted from population expansion. Mismatch analysis suggested that the expansion was initiated about 276,000 years ago for H. mohnikei and about 230,000 years ago for H. trimaculatus during the middle Pleistocene period. This study indicates a possible signature of genetic variation and population expansion in two seahorses under complex marine environments.

Proteomic identification of responsive proteins of Vibrio parahaemolyticus under high hydrostatic pressure.

BACKGROUND: High hydrostatic pressure (HHP) processing is currently being used as a treatment for certain foods to inhibit spoilage organisms and control the presence of foodborne pathogens. In this study proteome profiles were performed by two-dimensional gel electrophoresis (2-DE) coupled with MALDI-TOF/TOF identification to determine the effects of HHP (50, 100, 150 and 200 MPa, each for 10 min) on Vibrio parahaemolyticus ATCC 17802 (∼8 log CFU mL⁻¹) in order to understand how it responds to mechanical stress injury. RESULTS: Multiple comparisons of 2-DE revealed that the majority of changes in protein abundance occurred in a pressure-dependent fashion. A total of 18 differentially expressed protein spots were successfully identified by MALDI-TOF/TOF analysis. Moreover, quantitative RT-PCR and immunoblotting also substantiated the changes of transcriptional and translational levels of representative proteins. CONCLUSIONS: Our results suggested that V. parahaemolyticus may respond to HHP treatment through suppression of membrane stability and functionality (PfaC, Alr2, MltA, PLA2 and PatH), depression of biosynthesis and cellular processes (NadB, PyrB and ArgB), decreased levels of transcription (RpoD) and translation (RpsA, RplJ and PheS), and effective activation of protein folding and stress-related elements (GroES, DnaK and GroEL). This study may provide insight into the nature of the cellular targets of high pressure and in high-pressure resistance mechanisms in V. parahaemolyticus.

Reproductive behavior, embryonic and early larval development of the red head goby, Elacatinus puncticulatus.

The goals of this study are to provide a technical foundation for the production of the red head goby Elacatinus puncticulatus by evaluating its reproductive behavior and its embryonic and early larval development. Five pairs were kept under controlled conditions for thirty days. Courtship behavior, spawning period and the number of eggs produced were recorded. For the evaluation of embryo development, eggs were sampled at 12, 18, 24, 48, 72, 96, 120, 144 and 168h post-fertilization(HPF). To test the influence of the incubation period on larval total length and height, eggs with six days (6D) of incubation and with seven days of incubation (7D) were subjected to flashlight illumination for 30min to induce larval hatching. Another experiment evaluated the difference in larval survival with three different diets: Euplotes sp. (EU); rotifers Brachionus rotundiformis and Brachionus plicatilis and Paramecium sp. (BP); plankton collected from the wild (WP). The males displayed a gray head and pale yellow and black body coloration. Females exhibited strong red and black colors until three days before spawning, which occurred at intervals of 7 to 10 days. The hatching rate was 98-99%. The larvae total mean lengths and heights were 3.05 and 2.95mm (p>0.05) and 0.37 and 0.48mm (p<0.05) for treatments 6D and 7D, respectively. However, both groups exhibited high mortality at 5 days post-hatch (DPH). No larvae from the EU group survived after 5 DPH. At 8 DPH, 4% survivorship was found in treatment BP and 2% in treatment WP.

Identification and characterization of extracellular cyclic dipeptides as quorum-sensing signal molecules from Shewanella baltica, the specific spoilage organism of Pseudosciaena crocea during 4 °C storage.

Quorum-sensing (QS) signaling molecules are able to mediate specific gene expression inside spoilage bacteria in response to population density and thus are implicated in food spoilage. In the present work, a total of 102 strains of spoilage bacteria were isolated from Pseudosciaena crocea at 4 °C storage, and of these, 60 strains were identified as Shewanella spp., and 48 strains (47.1%) were identified as S. baltica. In addition, the spoilage capabilities of three different S. baltica strains (00A, 00B, and 00C) were compared by total volatile base nitrogen (TVB-N) and sensory analysis (off-odors). Furthermore, four cyclic dipeptides (diketopiperazines, DKPs) that function as QS signal molecules were isolated and characterized from the extracellular metabolites of S. baltica 00C which had the strongest spoilage activity based on gas chromatography mass spectrometry (GC-MS). By supplementation of four synthesized DKPs, the spoilage capability of S. baltica could be significantly enhanced. So far, this was the first attempt to characterize DKPs as the signaling molecules in QS of S. baltica. Our study may provide some evidence of the role of DKPs involved in microbial spoilage.

The dynamics of reproductive rate, offspring survivorship and growth in the lined seahorse, Hippocampus erectus Perry, 1810.

Seahorses are the vertebrate group with the embryonic development occurring within a special pouch in males. To understand the reproductive efficiency of the lined seahorse, Hippocampus erectus Perry, 1810 under controlled breeding experiments, we investigated the dynamics of reproductive rate, offspring survivorship and growth over births by the same male seahorses. The mean brood size of the 1-year old pairs in the 1(st) birth was 85.4±56.9 per brood, which was significantly smaller than that in the 6(th) birth (465.9±136.4 per brood) (P<0.001). The offspring survivorship and growth rate increased with the births. The fecundity was positively correlated with the length of brood pouches of males and trunk of females. The fecundity of 1-year old male and 2-year old female pairs was significantly higher than that from 1-year old couples (P<0.001). The brood size (552.7±150.4) of the males who mated with females that were isolated for the gamete-preparation, was larger than those (467.8±141.2) from the long-term pairs (P<0.05). Moreover, the offspring from the isolated females had higher survival and growth rates. Our results showed that the potential reproductive rate of seahorses H. erectus increased with the brood pouch development.

Effect of ferric ion on nitrogen consumption, biomass and oil accumulation of a Scenedesmus rubescens-like microalga.

This investigation examined the effects of ferric ion source and concentration on the biomass yield, fatty acid methyl esters (FAME) productivity and composition from Scenedesmus rubescens-like microalga. The algae cultivated with Fe(2+)-SO(4)(2-)/EDTA had higher ash free dry biomass (AFDB) production and in vivo chlorophyll a concentration than those with Fe(2+)-SO(4)(2-) and Fe(3+)-Cl(-) (P<0.001). The high or low Fe(2+) concentration can partly restrict the ammonia consumption, the AFDB and FAME accumulation. The algae at 1mg/L Fe(2+) concentration had the highest AFDB and FAME productivity (341.1 ± 25.8 and 107.8 ± 19.3mg/L/d) (P<0.001). The mean content of C16 and C18 series at all Fe(2+) concentrations was 88.6% ± 2.9% of total FAME, and it decreased with the increase of Fe(2+) concentrations. Among the fatty acids profiles, the contents of C18: 0, C18: 1n-7 and C18: 1n-9 are significantly changed under the different Fe(2+) concentrations, but not the contents of C16 series.

Effects of inorganic carbon concentration on carbon formation, nitrate utilization, biomass and oil accumulation of Nannochloropsis oculata CS 179.

This investigation examined the effects of the inorganic carbon concentration (4, 0.8 and 0 g/L NaHCO(3)) on the carbon formation, nitrate utilization, growth and fatty acids compositions of Nannochloropsis oculata. The dissolved inorganic carbon (DIC) concentration in the three treatments decreased sharply during the first 6 days, and the percentage of dissolved organic carbon (DOC) (% of total organic carbon (TOC)) decreased with the depletion of the DIC. The NO(3)(-) assimilation of the algae was correlated with the DIC concentration. The algae in the highest DIC treatment had the highest specific grow rate (0.0843 d(-1)) (P<0.0001), and their biomass and fatty acid methyl esters (FAME) productivity were 84.00 and 9.69 mg/L/d, respectively (P<0.0001). Contents of C16 and C18 series (% of FAME) were high and the C16:0 increased with the decrease of C18:1 during the cultivation. The iodine value (IV) of the algae was low at the low DIC media.

Effect of ultraviolet radiation on photosynthesis, biomass, and fatty acid content and profile of a Scenedesmus rubescens-like microalga.

The effects of ultraviolet radiation (UVR) and nitrogen (NaNO(3)) concentration on photosynthesis, biomass, and fatty acid content and profile of a Scenedesmus rubescens-like microalga were measured in an outdoor 8-day culture study. UV-induced photoinhibition decreased from 42.6% to 3.5%, in the presence of 75 mg/L NaNO(3) (HN) and from 52.9% to 22.6% in the presence of 7.5mg/L NaNO(3) (LN) nitrogen concentration, respectively. The concentrations of UV-absorbing compounds increased 4.3 and 4.9 times under HN and LN, respectively. Biomass accumulation was suppressed (10.7%) by UVR under HN, but not under LN conditions. Carotenoid content decreased from 1.05 ± 0.06 to 0.96 ± 0.15 (with UV radiation) and to 0.91 ± 0.07 (without UV radiation), respectively, under HN, while it decreased to 0.05 ± 0.04 (with UV radiation) and to 0.11 ± 0.08 (without UV radiation), respectively, under LN. The content of C18:1n9 fatty acids increased by about 430%, whereas that of C18:3n3 decreased by about 65% in both radiation treatments during nitrogen starvation. The results showed that the absence of UVR screening does not change the fatty acid content and profile of S. rubescens-like algae cultivated outdoors under HN and LN conditions.

Higher efficiency of CO2 injection into seawater by a venturi than a conventional diffuser system.

Mass production of microalgae generally requires the injection of CO(2) into open ponds or photo-bioreactors. The present study compares the CO(2) injection efficiency into seawater of a porous stone air diffuser and a venturi. CO(2) was injected at flow rates of 400, 700 and 1000 standard mL/min and 4, 7 and 10 standard L/min into a small and a large pond, respectively until the pH decreased from 7.8 to 6.8. No significant differences in CO(2) injection efficiency between the three CO(2) flow rates (p>0.05) were observed; however, CO(2) injection efficiency with venturi was about 100% (p<0.05) higher than that of the air diffuser. Therefore, it is possible to both reduce the cost and increase the effectiveness of CO(2) dissolution in seawater by using venturi operated at a lower flow rate, i.e. 400 standard mL/min in a small pond and 4 standard L/min in a large pond.

Polybrominated diphenyl ethers in water, sediment, soil, and biological samples from different industrial areas in Zhejiang, China.

Polybrominated diphenyl ethers (PBDEs) have been used extensively in electrical and electronic products, but little is known about their distribution in the environment surrounding the manufacturing factories. This study reports PBDE contamination in various matrices from the location (Liushi, Zhejiang province) that produces more than 70% of the low-voltage electrical appliances in China. Additionally, PBDE contamination was compared with other industries such as the e-waste recycling business (Fengjiang) in the same region. Specifically, we measured seven PBDE congeners (BDEs - 47, 99, 100, 153, 154, 183, and 209) in water, sediment, soil, plant, and animal tissues from four different areas in this region. The present study revealed elevated PBDE concentrations in all matrices collected from Liushi and Fengjiang in comparison with highly industrialized areas without significant PBDE contamination sources. In water samples, there were large variations of PBDE content and composition across different areas. In sediment/soil and biological samples, BDE-209 was the predominant congener and this could be due to the abundant usage of deca-BDE mixtures in China. Our findings provide the very first data on PBDE contamination in the local environments surrounding the electronics industry, and also reveal widespread PBDE contamination in highly industrialized coastal regions of China.

Biomass production and fatty acid profile of a Scenedesmus rubescens-like microalga.

This investigation examined the effects of nitrogen-phosphate combined deficiency on the biomass yield, fatty acid methyl esters (FAME) production and composition from Scenedesmus rubescens-like microalga. A 15-day indoor culture was performed as a 3 × 3 factorial design (NaNO(3) levels: 3, 10 and 20mM; KH(2)PO(4) levels: 20, 50 and 150 µM). The algae grown under medium nitrogen concentration (10mM) and high phosphate concentration (150 µM) reached the highest biomass (1223.5 ± 152.5mg/L). Both nitrogen and phosphate had a significant influence on the FAME yield (P<0.05 and P<0.0001, respectively). The FAME yield from algae grown under low nitrogen (3mM) and phosphate concentration (20 µM) increased throughout the experiment and the highest FAME yield (42.2 ± 2.5% of AFDW) as well as C16 and C18 content (95.8 ± 1.6% of AFDW) was achieved under these conditions. Algae grown under medium nitrogen concentration (10mM) and low phosphate concentration (20 µM) had the highest FAME productivity (426.0mg/L ± 135.0mg/L). Thus, the lower nitrogen concentration (3mM-10mM) and low phosphate concentration (20 µM) would be an optimal combination tested to produce the most FAME from S. rubescens-like algae.

Simultaneously hermaphroditic shrimp use lipophilic cuticular hydrocarbons as contact sex pheromones.

Successful mating is essentially a consequence of making the right choices at the correct time. Animals use specific strategies to gain information about a potential mate, which is then applied to decision-making processes. Amongst the many informative signals, odor cues such as sex pheromones play important ecological roles in coordinating mating behavior, enabling mate and kin recognition, qualifying mate choice, and preventing gene exchange among individuals from different populations and species. Despite overwhelming behavioral evidence, the chemical identity of most cues used in aquatic organisms remains unknown and their impact and omnipresence have not been fully recognized. In many crustaceans, including lobsters and shrimps, reproduction happens through a cascade of events ranging from initial attraction to formation of a mating pair eventually leading to mating. We examined the hypothesis that contact pheromones on the female body surface of the hermaphroditic shrimp Lysmata boggessi are of lipophilic nature, and resemble insect cuticular hydrocarbon contact cues. Via chemical analyses and behavioural assays, we show that newly molted euhermaphrodite-phase shrimp contain a bouquet of odor compounds. Of these, (Z)-9-octadecenamide is the key odor with hexadecanamide and methyl linoleate enhancing the bioactivity of the pheromone blend. Our results show that in aquatic systems lipophilic, cuticular hydrocarbon contact sex pheromones exist; this raises questions on how hydrocarbon contact signals evolved and how widespread these are in the marine environment.

Effects of nitrogen source and concentration on biomass and oil production of a Scenedesmus rubescens like microalga.

This investigation examined the effects of nitrogen source and concentration on the ash free dry biomass (AFDB) and oil accumulation and productivity of a Scenedesmusrubescens like alga cultivated in indoor photo-bioreactors and outdoor raceway ponds. Among the nitrogen sources ((NH4)2CO3, urea, NaNO3, urea and NaNO3 mixture), the algae fed with the mixture of urea-N and NaNO3-N had the highest AFDB productivity of 0.539±0.040 g/L/d and the content of fatty acid methyl esters (FAME) (%) fed with (NH4)2CO3-N increased continuously for 17 days and reached 42.94±2.05% in the indoor photo-bioreactors. The content of C18 series (as % of total FAME) was high in all the treatments of nitrogen sources. The algae fed with (NH4)2CO3-N had the highest FAME productivity of 0.133 g/L/d (P<0.001). The effect of ammonia concentration on the AFDB and FAME productivity was also studied during the 20-day cultivation in the outdoor raceway cultivation system. The mean FAME of the algae cultivated in the ammonia concentration of 1.0-1.5 mg/L were the highest (0.556±0.112 g/m2/d) (P<0.001). The total Kjeldahl nitrogen (TKN) in the algae and cultivation water decreased but the FAME content (%) of the algae increased during the nitrogen starvation phase.

Aquatic organotin pollution in Taiwan.

The current status of aquatic organotin pollution in Taiwan is reviewed. In freshwater sediments and biota, especially in rice-field related habitats, phenyltins (PTs) were dominant among the organotin pollutants, whereas butyltins (BTs) were usually predominant in marine environments. Among the marine habitats, contamination levels were found to be in the descending order of harbour and estuary>fish cultural site>coastal>offshore>coral reefs. Imposex snails were observed in all the sampling years (1990-2003). Meanwhile, organotin concentrations were greater in winter than those in summer, whereas proportions of PTs were much higher in summer than in winter. Due to the lack of continuous monitoring data, the effectiveness of the ban on TPT usage in agriculture in 1999 and the prohibition of TBT use on small boats in 2003 is still not known.