Prospective machine learning CT quantitative evaluation of idiopathic pulmonary fibrosis in patients undergoing anti-fibrotic treatment using low- and ultra-low-dose CT.

AIM: To compare the machine learning computed tomography (CT) quantification tool, Computer-Aided Lung Informatics for Pathology Evaluation and Ratings (CALIPER) to pulmonary function testing (PFT) in assessing idiopathic pulmonary fibrosis (IPF) for patients undergoing treatment and determine the effects of limited (LD) and ultra-low dose (ULD) CT on CALIPER performance. MATERIALS AND METHODS: Thirty-eight IPF patients underwent PFT and standard, LD, and ULD CT. CALIPER classified each CT voxel into either vessel-related structures (VRS), normal, reticular (R), honeycomb (HC) or ground-glass (GG) features. CALIPER-derived interstitial lung disease (ILD) extent represented the sum of GG, R and HC values. Repeated-measures correlation coefficient (ρrm) and 95% confidence interval (CI) evaluated CALIPER features correlation with PFT. Lin's concordance correlation coefficient (CCC) assessed concordance of CALIPER parameters across different CT dosages. RESULTS: Twenty patients completed 12 months of follow-up. CALIPER ILD correlated significantly with percent predicted (%) forced vital capacity (FVC) and forced expiratory volume in 1 second (%FEV1; p=0.004, ρrm -0.343, 95% CI [-0.547, -0.108] and 0.008, -0.321, [-0.518, -0.07], respectively). VRS significantly correlated with %FVC and %FEV1 (p=0.000, ρrm -0.491, 95% CI [-0.685, -0.251] and -0.478, 0.000, [-0.653, -0.231], respectively). There was near perfect LD and moderate ULD concordance with standard dose CT for both ILD (CCC 0.995, 95% CI 0.988-0.999 and 0.9, 0.795-0.983, respectively) and VRS (CCC 0.989, 95% CI 0.963-0.997 and 0.915, 0.806-0.956, respectively). CONCLUSIONS: CALIPER parameters correlate well with PFTs for evaluation of IPF in patients undergoing anti-fibrotic treatment without being influenced by dose variation. CALIPER may serve as a robust, objective adjunct to PFTs in assessing anti-fibrotic treatment related changes.

Molecular identification, antifungal susceptibility profile, and biofilm formation of clinical and environmental Rhodotorula species isolates.

Rhodotorula species are emergent fungal pathogens capable of causing invasive infections, primarily fungemia. They are particularly problematic in immunosuppressed patients when using a central venous catheter. In this study, we evaluated the species distribution of 51 clinical and 8 environmental Rhodotorula species isolates using the ID32C system and internal transcribed spacer (ITS) sequencing. Antifungal susceptibility testing and biofilm formation capability using a crystal violet staining assay were performed. Using ITS sequencing as the gold standard, the clinical isolates were identified as follows: 44 R. mucilaginosa isolates, 2 R. glutinis isolates, 2 R. minuta isolates, 2 R. dairenensis isolates, and 1 Rhodosporidium fluviale isolate. The environmental isolates included 7 R. mucilaginosa isolates and 1 R. slooffiae isolate. Using the ID32C system, along with a nitrate assimilation test, only 90.3% of the isolates tested were correctly identified. In the biofilm formation assay, R. mucilaginosa and R. minuta exhibited greater biofilm formation ability compared to the other Rhodotorula species; the clinical isolates of R. mucilaginosa showed greater biofilm formation compared to the environmental isolates (P = 0.04). Amphotericin B showed good in vitro activity (MIC ≤ 1 µg/ml) against planktonic cells, whereas voriconazole and posaconazole showed poor activity (MIC(50)/MIC(90), 2/4 µg/ml). Caspofungin and fluconazole MICs were consistently high for all isolates tested (≥64 µg/ml and ≥ 4 µg/ml, respectively). In this study, we emphasized the importance of molecular methods to correctly identify Rhodotorula species isolates and non-R. mucilaginosa species in particular. The antifungal susceptibility profile reinforces amphotericin B as the antifungal drug of choice for the treatment of Rhodotorula infections. To our knowledge, this is the first study evaluating putative differences in the ability of biofilm formation among different Rhodotorula species.

Coverage path planning for spraying drones.

The pandemic by COVID-19 is causing a devastating effect on the health of the global population. Currently, there are several efforts to prevent the spread of the virus. Among those efforts, cleaning and disinfecting public areas have become important tasks and they should be automated in future smart cities. To contribute in this direction, this paper proposes a coverage path planning method for a spraying drone, an unmanned aerial vehicle that has mounted a sprayer/sprinkler system, that can disinfect areas. State-of-the-art planners consider a camera instead of a sprinkler, in consequence, the expected coverage will differ in running time because the liquid dispersion is different from a camera's projection model. In addition, current planners assume that the vehicles can fly outside the target region; this assumption can not be satisfied in our problem, because disinfections are performed at low altitudes. Our method presents i) a new sprayer/sprinkler model that fits a more realistic coverage volume to the drop dispersion and ii) a planning method that efficiently restricts the flight to the region of interest avoiding potential collisions in bounded scenes. The algorithm has been tested in several simulation scenes, showing that it is effective and covers more areas with respect to two approaches in the literature. Note that the proposal is not limited to disinfection applications, but can be applied to other ones, such as painting or precision agriculture.

Comparison of [¹¹¹In]pentetreotide-SPECT and [¹8F]FDOPA-PET in the localization of extra-adrenal paragangliomas: the case for a patient-tailored use of nuclear imaging modalities.

AIMS AND METHODS: The aim of this prospective study was to compare the diagnostic value of [¹8F]FDOPA-PET and [¹¹¹In]pentetreotide-SPECT somatostatin receptor scintigraphy (SRS) in patients with nonmetastatic extra-adrenal paragangliomas (PGLs). Twenty-five consecutive unrelated patients who were known or suspected of having nonmetastatic extra-adrenal PGLs were prospectively evaluated with SRS and [¹8F]FDOPA-PET. ¹³¹I-MIBG and [¹8F]FDG-PET were added to the work-up in patients with a personal or familial history of PGL, predisposing mutations, abdominal PGLs, metanephrine hypersecretion and abdominal foci on SRS and/or [¹8F]FDOPA-PET. RESULTS: SRS correctly detected 23/45 lesions of which 20 were head or neck lesions (H&N) and 3 were abdominal lesions. [¹8F]FDOPA-PET detected significantly more lesions than SRS (39/45, P < 0·001). Both SRS and ¹8F-DOPA-PET detected significantly more H&N than abdominal lesions (66·7% vs 20%, P = 0·003 and 96·7% vs 67%, P = 0·012, respectively). In two patients with the succinate dehydrogenase D (SDHD) mutation, [¹8F]FDOPA-PET missed five abdominal PGLs which were detected by the combination of SRS, [¹³¹I]MIBG and [¹8F]FDG-PET. A lesion-based analysis using a forward stepwise logistic regression model demonstrates that size ≤ 10 mm (P = 0·002) and abdominal lesions (P = 0·031) were independently associated with "[¹8F]FDOPA-PET diagnosis only". In turn, a previous history of surgery and/or the presence of germline mutation was associated with lower lesion size (P = 0·001). CONCLUSIONS: The sensitivity of SRS for localizing parasympathetic PGLs is lower than originally reported, and [¹8F]FDOPA-PET is better than SRS for localizing small lesions. SRS should be replaced by [¹8F]FDOPA-PET as the first-line imaging procedure in H&N PGL, especially in patients at risk of multifocal disease (predisposing mutations and or previous history of surgery).

Pectin lyase from Aspergillus giganteus: comparative study of productivity of submerged fermentation on citrus pectin and orange waste.

The aim of this study was to investigate some of the factors affecting pectin lyase (PL) production by an Aspergillus giganteus strain, and to characterize this pectinolytic activity excreted into the medium. The highest activities were obtained with orange waste, citrus pectin and galacturonic acid as carbon sources. The highest activity, using citrus pectin as carbon source, was obtained in 11-day-old standing cultures, but the highest specific activity was obtained in 6.5-day-old shaken cultures, at pH 6.5 and 35 degrees C. Using orange waste as carbon source, the highest activity was observed in 8-day-old standing cultures, at pH 7.0 and 30 degrees C. Optimal assay conditions were pH 8.5-9.0 and 50 degrees C. The PL activity showed thermal stability, with half-lives of 30 and 27 min when incubated at 45 and 50 degrees C, respectively. High stability was observed at room temperature from pH 6.0 to 10.0; more than 85% of enzyme activity was preserved in this pH range. Under optimum conditions, the highest pectin lyase activity in the medium was 470 U/ml, with orange waste as carbon source.

Production of beta-galactosidase by Trichoderma reesei FTKO-39 in wheat bran: partial purification of two isozymes.

Trichoderma reesei FTKO-39 grown at 35 degrees C for 5 d on wheat bran supplemented with MgCl2 and lactose as the carbon source produced two isozymes of beta-galactosidase: BGT I and BGT II. These isozymes were partially purified on a DEAE-Trisacryl column. Both BGT I and BGT II fractions exhibited optimum activity at 65 degrees C, but the pH optima were 4.0 and 6.5, respectively. The isozymes also showed similar thermal stability. However, BGT I was more stable than BGT II in a pH range of 3.0-10.0. At least two different beta-galactosidases are produced by T. reesei, as revealed by the two bands seen on a 6% polyacrylamide gel stained for activity.

Micro-dissected tumor tissues on chip: an ex vivo method for drug testing and personalized therapy.

In cancer research and personalized medicine, new tissue culture models are needed to better predict the response of patients to therapies. With a concern for the small volume of tissue typically obtained through a biopsy, we describe a method to reproducibly section live tumor tissue to submillimeter sizes. These micro-dissected tissues (MDTs) share with spheroids the advantages of being easily manipulated on-chip and kept alive for periods extending over one week, while being biologically relevant for numerous assays. At dimensions below ~420 µm in diameter, as suggested by a simple metabolite transport model and confirmed experimentally, continuous perfusion is not required to keep samples alive, considerably simplifying the technical challenges. For the long-term culture of MDTs, we describe a simple microfluidic platform that can reliably trap samples in a low shear stress environment. We report the analysis of MDT viability for eight different types of tissues (four mouse xenografts derived from human cancer cell lines, three from ovarian and prostate cancer patients, and one from a patient with benign prostatic hyperplasia) analyzed by both confocal microscopy and flow cytometry over an 8-day incubation period. Finally, we provide a proof of principle for chemosensitivity testing of human tissue from a cancer patient performed using the described MDT chip method. This technology has the potential to improve treatment success rates by identifying potential responders earlier during the course of treatment and providing opportunities for direct drug testing on patient tissues in early drug development stages.

[Papillary carcinoma of the thyroid metastatic to the parapharyngeal space]. / Tumoración parafaríngea metastásica de carcinoma papilar de tiroides.

This is a case report of a papillary thyroid carcinoma metastatic to the parapharyngeal space, that presented with local obstructive symptoms (dysphagia and displacement of the ipsilateral tonsil). The diagnosis was suspected by imaging studies (CT and MRI) but not confirmed until histological examination. It is not common for such tumors to metastasize to the parapharyngeal space. This is the reason why, to our knowledge, there have been described only five similar cases previously.

Monitoring the Diversity of Hunting Ants (Hymenoptera: Formicidae) on a Fragmented and Restored Andean Landscape.

Hunting ants are predators of organisms belonging to different trophic levels. Their presence, abundance, and diversity may reflect the diversity of other ants and contribute to evaluate habitat conditions. Between 2003 and 2005 the restoration of seven corridors in an Andean rural landscape of Colombia was performed. The restoration took place in lands that were formerly either forestry plantations or pasturelands. To evaluate restoration progress, hunting ants were intensely sampled for 7 yr, using sifted leaf litter and mini-Winkler, and pitfall traps in 21 plots classified into five vegetation types: forests, riparian forests, two types of restored corridors, and pasturelands. The ant communities were faithful to their habitat over time, and the main differences in ant composition, abundance, and richness were due to differences among land use types. The forests and riparian forests support 45% of the species in the landscape while the restored corridors contain between 8.3-25%. The change from forest to pasturelands represents a loss of 80% of the species. Ant composition in restored corridors was significantly different than in forests but restored corridors of soil of forestry plantations retained 16.7% more species than restored corridors from pasturelands. Ubiquitous hunting ants, Hypoponera opacior (Forel) and Gnamptogenys ca andina were usually associated with pastures and dominate restored corridors. Other cryptic, small, and specialized hunting ants are not present in the restored corridors. Results suggest that the history of land use is important for the biodiversity of hunting ants but also that corridors have not yet effectively contributed toward conservation goals.

The specificity of the S1' subsite of cysteine proteases.

The specificity of the S1' subsite of the cysteine proteases cathepsin B, L, S and papain has been investigated using a series of intramolecularly quenched fluorogenic substrates (Dansyl-Phe-Arg-AA-Trp-Ala) where the P1' amino acid (AA) has been varied. Taken individually, each enzyme displays a relatively broad S1' subsite specificity and this subsite cannot be considered as a primary site of specificity. Notable differences do exist however between the various proteases. Cathepsin B prefers large hydrophobic residues in the P1' position of a substrate while cathepsin L has an opposite trend, favoring amino acids with small (Ala, Ser) or long but non-branched (Asn, Gln, Lys) side chains. Cathepsin S and papain display a somewhat broader S1' subsite specificity.

Inhibitors of the major cysteinyl proteinase (GP57/51) impair host cell invasion and arrest the intracellular development of Trypanosoma cruzi in vitro.

Peptidyl diazomethane (PDAM) derivatives, a class of irreversible inhibitors for cysteine proteinase, were screened for the ability to impair Trypanosoma cruzi invasion and intracellular development in primary cultures of heart muscle cells (HMC). T. cruzi GP57/51, a purified cysteinyl proteinase, and the substrate Z-Phe-Arg-NHMec were used to determine inhibition rate constants (k'+2) by continuous kinetic assays. The k'+2 values ranged from 25,400 to 2,800. The best inhibitors of GP57/51 had bulky hydrophobic residues in the P1 position (in addition to P2), the S1 sub-site specificity of the enzyme being thus similar to mammalian cathepsin L. The effects of these PDAM on parasite infectivity were then investigated. The ability to invade HMC was markedly impaired when trypomastigotes were briefly exposed to 10 microM of Z-(S-Bzl)Cys-Phe-CHN2. Striking effects were observed when PDAM were added to HMC cultures that had been previously infected with trypomastigotes: Z-(S-Bzl)Cys-Phe-CHN2 with an IC50 of 0.4 microM, and less markedly Z-Phe-Phe-CHN2 and Z-Tyr-Phe-CHN2 (or Z-Phe-Tyr-CHN2) blocked amastigote replication as well as their transformation into trypomastigotes, thereby arresting intracellular development. Bz-Phe-Gly-CHN2, in contrast, failed to display antiparasite activity. Direct characterization of the target cysteinyl proteinase was sought, by incubating viable amastigotes or infected HMC with Z-[125I]Tyr-Phe-CHN2. Affinity labeling implicated GP57/51 as the major cysteinyl proteinase target for this probe. We propose that T. cruzi intracellular development is critically dependent on GP57/51 (cruzipain). Selective inhibitors for this cysteinyl proteinase may have therapeutic potential.

Ribosomal RNA sequences of Sarcocystis muris, Theileria annulata and Crypthecodinium cohnii reveal evolutionary relationships among apicomplexans, dinoflagellates, and ciliates.

Sarcocystis muris is a coccidium with a two-host life cycle involving the domestic cat and the mouse, Mus musculus. S. muris and Theileria annulata belong to the phylum Apicomplexa, but the latter organism is a tick-borne protozoon in the subclass Piroplasmea and causes tropical theileriosis in cattle. The small-subunit ribosomal RNA (16S-like rRNA) coding regions of these organisms as well as that of the free living dinoflagellate Crypthecodinium cohnii were amplified using polymerase chain reaction techniques and compared to 16S-like rRNA sequences from other eukaryotes. The 16S-like rRNA genes of S. muris and T. annulata are more similar to each other than either is to Plasmodium falciparum, the cause of malignant tertian malaria of humans or Plasmodium berghei, the agent of the commonly studied malaria of rodents. Evolutionary trees inferred from the rRNA sequence similarities support a close phylogenetic relationship between the Apicomplexa and Dinoflagellata as represented by Prorocentrum micans and C. cohnii. Apparently members of these related phyla arose from an ancestral stock that gave rise to the ciliated protozoa.

A standardized method for measuring anti-F VIII: C inhibitors in haemophilia A by coagulation inhibition in agarose gel.

Antibodies against factor-VIII coagulant activity can appear in haemophilic patients and, although infrequently, can affect individuals not suffering from Haemophilia A. The Oxford and Bethesda methods are presently the most commonly used techniques for measuring these antibodies. Both methods are time-consuming and not suitable for the screening of large risk groups. An appropriate method for screening these coagulation inhibitors is that described by P. Bird in 1975. It is based on inhibition of the coagulation produced when plasma samples containing inhibitors diffuse in agarose gels mixed with normal platelet rich plasma (PRP). However, this technique is highly dependent on the variability derived from the use of PRP (amount of coagulant factor-VIII, number of platelets, etc.). In an attempt to avoid these disadvantages, Bird's method has been modified by using standardized commercial reagents (lyophilized plasma with 100% factor-VIII coagulant activity, purified fibrinogen, and platelet Factor 3) instead of PRP. The sensitivity reaches 0.8 Bethesda units and the correlation with the Bethesda method is r = 0.964, p less than 0.001. This newly developed method is as simple as Bird's, and appears to be at least, as accurate and reproducible as the Bethesda method.

The pagetoid variant of bladder urothelial carcinoma in situ A clinicopathological study of 11 cases.

Pagetoid urothelial carcinoma in situ (CIS) is a rare variant of bladder cancer that is characterized by an intraepithelial proliferation of large cells arranged singly or in clusters and randomly distributed. These neoplasms deserve recognition and attention, chiefly because they may be overlooked or misdiagnosed as urothelial dysplasia, then causing unsuspected tumor recurrence after surgery. We report on the clinicopathological features and immunohistochemical findings of 11 (14.86%) cases of pagetoid CIS in a retrospective study of 74 cases of conventional carcinoma in situ. Most patients were male ( n=10). Their ages ranged from 31 years to 78 years. The lesion can be present with primary ( n=2) or secondary ( n=9) CIS. Pagetoid CIS is usually a focal lesion occurring in a clinical and histological setting of conventional CIS, and these patients essentially have the same progression and survival rates as patients without pagetoid changes and are treated in the same way. In cases with extensive urothelial denudation, pagetoid CIS may be focally present in otherwise normal-looking urothelium, thus alerting the pathologist to search for additional CIS elsewhere in the bladder. Given that primary extramammary Paget disease of the external genitalia and of the anal canal may extend to the bladder and, conversely, some bladder cases of pagetoid CIS may extend to the urethra, ureter, and beyond to the external genitalia, the differential diagnoses between these two entities represent an important therapeutic consideration. Our data suggest that a panel of immunostains including CK7+/CK20+/TM+ may assist in differentiating urothelial pagetoid CIS from extramammary Paget disease which is known to be CK7+/CK20-.

Synthesis and structural characterization of Be(eta 5-C5Me5)(eta 1-C5Me4H). Evidence for ring-inversion leading to Be(eta 5-C5Me4H)(eta 1-C5Me5).

The mixed-ring beryllocene Be(C5Me5)(C5Me4H), that contains eta 5-C5Me5 and eta 1-C5Me4H rings, the latter bonded to the metal through the CH carbon atom (X-ray crystal structure) reacts at room temperature with CNXyl (Xyl = C6H3-2,6-Me2) to give an iminoacyl product, Be(eta 5-C5Me4H)[C(NXyl)C5Me5] derived from the inverted beryllocene structure Be (eta 5-C5Me4H)(eta 1-C5Me5).

Detection of two different anti-factor VIII/von Willebrand factor antibodies of the IgA class in a hemophilic patient with a polyclonal factor VIII inhibitor of the IgG class.

A hemophilic patient treated with Factor VIII (F.VIII) concentrates developed a F.VIII inhibitory activity. The patient's plasma showed 2 anti-F.VIII/von Willebrand Factor (vWF) antibodies (Abs) of the IgG and IgA class respectively. The specific anti-F.VIII/vWF Abs were isolated and resulted to be IgG (20 micrograms/ml) and IgA (35 micrograms/ml). The plasma was subsequently fractionated by Protein A-Sepharose and the IgG and IgA containing fractions were separately analyzed for anti-F.VIII activity. Both fractions exhibited F.VIII inhibitory activity, but that corresponding to the IgA was lower than expected. Regarding the possible existence of a non-inhibitor Ab in the IgA containing sample, plasma was processed through an immunoadsorbent to which a F.VIII devoid vWF preparation had been previously coupled and further fractionated by Protein A-Sepharose, obtaining 2 IgA fractions. One of them exhibited F.VIII inhibitory activity while the other, which reacted with the F.VIII devoid vWF preparation, did not. Therefore, this latter one was considered as a true anti-vWF Ab. The IgG and IgA inhibitors were polyclonal and the IgG one was composed of the 4 IgG subclasses.

Immunological characterization of factor VIII inhibitors by a sensitive micro-ELISA method.

A micro-ELISA method for the immunological characterization of factor VIII (F. VIII) inhibitors is described. Microplates sensitized with purified monospecific anti-von Willebrand factor (vWF) IgG were firstly incubated with a commercial F. VIII concentrate and then with the samples containing the F. VIII inhibitors to be characterized. Peroxidase conjugated monospecific antisera were used to determine the Ig class and the light chain type. Alkaline phosphatase conjugated monoclonal antibodies were employed to investigate the IgG subclasses. The F. VIII inhibitors from 8 hemophiliacs and 1 patient with systemic lupus erythematosus (SLE) were characterized. All of them belonged to the IgG class but one case simultaneously exhibited another inhibitor of the IgA class. Eight inhibitors analyzed for light chains resulted to be polyclonal. In 7 cases the inhibitors belonged solely to the IgG4 subclass while in the other 2 cases contained all the 4 IgG subclasses. The method appears to be simple, accurate and highly sensitive (detecting as low as 0.156 Bethesda Units/ml) for the immunological characterization of F. VIII inhibitors.

Properties of a polynucleotide synthesized by strain 74A of Neurospora crassa.

A polynucleotide (or a fragment of RNA) was purified to apparent homogeneity by HPLC from mycelium of the wild strain 74A of the mould Neurospora crassa, after growth on sucrose and in the presence of saturating amounts of inorganic phosphate (Pi) for 72 hr at 30 degrees. The M(r) was ca 20,000 as determined by HPLC at pH 6.8. Polynucleotide synthesis ranged from 4.0 to 6.5 micrograms polynucleotide per mg dry mycelium in mycelium of the wild strain 74A and the various phosphorus regulatory and structural mutant strains of the mould N. crassa. Kinetic data showed that the polynucleotide interacts with mycelial Pi-repressible alkaline phosphatase by inhibiting its p-nitrophenylphosphatase activity and by protecting the enzyme against thermal inactivation in the presence of high concentrations of ammonium sulphate.

On the importance of GABA-ergic neurons for the AOAA induced accumulation of GABA in the rat brain.

The accumulation of GABA induced by an intravenous injection of aminooxyacetic acid (AOAA) was followed for 1 h in five parts of the rat brain, i.e. cerebellum, medulla oblongata-pons, striatum, ventral mesencephalon and hypothalamus. The accumulation was similar in all brain parts studied when expressed as percentual increase from the basal value; an initial rapid accumulation indicating a mean turnovertime of 12--16 min during the first 5 min was gradually decreased to turnovertime of about 1--3 h during the last 30 min of observation. In order experiments brains were transected unilaterally between the substantia nigra and the striatum. Six days after the hemisection the GABA concentration of the substantia nigra of the transected side had decreased to less than 30% of the control side. The AOAA induced accumulation of GABA in the substantia nigra of the transected side was less pronounced than that of the control side. The initial rapid accumulation seen in all brain parts studied was completely lacking in the substantia nigra of the transected side. In the striatum, the transection did neither alter the GABA concentration nor the AOAA induced accumulation of GABA. As the initial rapid accumulation of GABA disappears after degeneration of GABA-ergic neurons, it is suggested that this initial phase of GABA accumulation produced by an intravenous injection of AOAA probably is the result of GABA accumulation in neurons.