Establishment of the deuterium oxide dilution method as a new possibility for determining the transendothelial water permeability.

Increase in transendothelial water permeability is an essential etiological factor in a variety of diseases like edema and shock. Despite the high clinical relevance, there has been no precise method to detect transendothelial water flow until now. The deuterium oxide (D2O) dilution method, already established for measuring transepithelial water transport, was used to precisely determine the transendothelial water permeability. It detected appropriate transendothelial water flow induced by different hydrostatic forces. This was shown in four different endothelial cell types. The general experimental setup was verified by gravimetry and absorbance spectroscopy. Determination of transendothelial electrical resistance (TEER) and immunocytochemical staining for proteins of the cell-cell contacts were performed to ensure that no damage to the endothelium occurred because of the measurements. Furthermore, endothelial barrier function was modulated. Measurement of transendothelial water flux was verified by measuring the TEER, the apparent permeability coefficient and the electrical capacity. The barrier-promoting substances cyclic adenosine monophosphate and iloprost reduced TEER and electrical capacity and increased permeability. This was accompanied by a reduced transendothelial water flux. In contrast, the barrier-damaging substances thrombin, histamine and bradykinin reduced TEER and electrical capacity, but increased permeability. Here, an increased water flow was shown. This newly established in vitro method for direct measurement of transendothelial water permeability was verified as a highly precise technique in various assays. The use of patient-specific endothelial cells enables individualized precision medicine in the context of basic edema research, for example regarding the development of barrier-protective pharmaceuticals.

Conceptualizing flexible papers using cellulose model surfaces and polymer particles.

Cellulose, as a naturally abundant and biocompatible material, is still gaining interest due to its high potential for functionalization. This makes cellulose a promising candidate for replacing plastics. Understanding how cellulose interacts with various additives is crucial for creating composite materials with diverse properties, as it is the case for plastics. In addition, the mechanical properties of the composite materials are assumed to be related to the mobility of the additives against the cellulose. Using a well-defined cellulose model surface (CMS), we aim to understand the adsorption and desorption of two polymeric particles (core-shell particles and microgels) to/from the cellulose surface. The nanomechanics of particles and CMS are quantified by indentation measurements with an atomic force microscope (AFM). AFM topography measurements quantified particle adsorption and desorption on the CMS, while peak force AFM measurements determined the force needed to move individual particles. Both particles and the CMS exhibited pH-dependent charge behavior, allowing a tunable interaction between them. Particle adsorption was irreversible and driven by electrostatic forces. In contrast, desorption and particle mobility forces are dominated by structural morphology. In addition, we found that an annealing procedure consisting of swelling/drying cycles significantly increased the adhesion strength of both particles. Using the data, we achieve a deeper understanding of the interaction of cellulose with polymeric particles, with the potential to advance the development of functional materials and contribute to various fields, including smart packaging, sensors, and biomedical applications.

The cyto-linker and scaffolding protein "plectin" mis-localization leads to softening of cancer cells.

Discovering tools to prevent cancer progression requires understanding the fundamental differences between normal and cancer cells. More than a decade ago, atomic force microscopy (AFM) revealed cancer cells' softer body compared to their healthy counterparts. Here, we investigated the mechanism underlying the softening of cancerous cells in comparison with their healthy counterparts based on AFM high resolution stiffness tomography and 3D confocal microscopy. We showed microtubules (MTs) network in invasive ductal carcinoma cell cytoskeleton is basally located and segmented for around 400 nm from the cell periphery. Additionally, the cytoskeleton scaffolding protein plectin exhibits a mis-localization from the cytoplasm to the surface of cells in the carcinoma which justifies the dissociation of the MT network from the cell's cortex. Furthermore, the assessment of MTs' persistence length using a worm-like-chain (WLC) model in high resolution AFM images showed lower persistence length of the single MTs in ductal carcinoma compared to that in the normal state. Overall, these tuned mechanics support the invasive cells to ascertain more flexibility under compressive forces in small deformations. These data provide new insights into the structural origins of cancer aids in progression.

Self-Assembly of Polymer-Modified FePt Magnetic Nanoparticles and Block Copolymers.

The fabrication of nanocomposites containing magnetic nanoparticles is gaining interest as a model for application in small electronic devices. The self-assembly of block copolymers (BCPs) makes these materials ideal for use as a soft matrix to support the structural ordering of the nanoparticles. In this work, a high-molecular-weight polystyrene-b-poly(methyl methacrylate) block copolymer (PS-b-PMMA) was synthesized through anionic polymerization. The influence of the addition of different ratios of PMMA-coated FePt nanoparticles (NPs) on the self-assembled morphology was investigated using transmission electron microscopy (TEM) and small-angle X-ray scattering (SAXS). The self-assembly of the NPs inside the PMMA phase at low particle concentrations was analyzed statistically, and the negative effect of higher particle ratios on the lamellar BCP morphology became visible. The placement of the NPs inside the PMMA phase was also compared to theoretical descriptions. The magnetic addressability of the FePt nanoparticles inside the nanocomposite films was finally analyzed using bimodal magnetic force microscopy and proved the magnetic nature of the nanoparticles inside the microphase-separated BCP films.

Microscopic Evidence of Malaria Infection in Visceral Tissue from Medici Family, Italy.

Microscopy of mummified visceral tissue from a Medici family member in Italy identified a potential blood vessel containing erythrocytes. Giemsa staining, atomic force microscopy, and immunohistochemistry confirmed Plasmodium falciparum inside those erythrocytes. Our results indicate an ancient Mediterranean presence of P. falciparum, which remains responsible for most malaria deaths in Africa.

Differential cortical activation patterns: pioneering sub-classification of tinnitus with and without hyperacusis by combining audiometry, gamma oscillations, and hemodynamics.

The ongoing controversies about the neural basis of tinnitus, whether linked with central neural gain or not, may hamper efforts to develop therapies. We asked to what extent measurable audiometric characteristics of tinnitus without (T) or with co-occurrence of hyperacusis (TH) are distinguishable on the level of cortical responses. To accomplish this, electroencephalography (EEG) and concurrent functional near-infrared spectroscopy (fNIRS) were measured while patients performed an attentionally demanding auditory discrimination task using stimuli within the individual tinnitus frequency (fTin) and a reference frequency (fRef). Resting-state-fMRI-based functional connectivity (rs-fMRI-bfc) in ascending auditory nuclei (AAN), the primary auditory cortex (AC-I), and four other regions relevant for directing attention or regulating distress in temporal, parietal, and prefrontal cortex was compiled and compared to EEG and concurrent fNIRS activity in the same brain areas. We observed no group differences in pure-tone audiometry (PTA) between 10 and 16 kHz. However, the PTA threshold around the tinnitus pitch was positively correlated with the self-rated tinnitus loudness and also correlated with distress in T-groups, while TH experienced their tinnitus loudness at minimal loudness levels already with maximal suffering scores. The T-group exhibited prolonged auditory brain stem (ABR) wave I latency and reduced ABR wave V amplitudes (indicating reduced neural synchrony in the brainstem), which were associated with lower rs-fMRI-bfc between AAN and the AC-I, as observed in previous studies. In T-subjects, these features were linked with elevated spontaneous and reduced evoked gamma oscillations and with reduced deoxygenated hemoglobin (deoxy-Hb) concentrations in response to stimulation with lower frequencies in temporal cortex (Brodmann area (BA) 41, 42, 22), implying less synchronous auditory responses during active auditory discrimination of reference frequencies. In contrast, in the TH-group gamma oscillations and hemodynamic responses in temporoparietal regions were reversed during active discrimination of tinnitus frequencies. Our findings suggest that T and TH differ in auditory discrimination and memory-dependent directed attention during active discrimination at either tinnitus or reference frequencies, offering a test paradigm that may allow for more precise sub-classification of tinnitus and future improved treatment approaches.

Concentration gradients in evaporating binary droplets probed by spatially resolved Raman and NMR spectroscopy.

Understanding the evaporation process of binary sessile droplets is essential for optimizing various technical processes, such as inkjet printing or heat transfer. Liquid mixtures whose evaporation and wetting properties may differ significantly from those of pure liquids are particularly interesting. Concentration gradients may occur in these binary droplets. The challenge is to measure concentration gradients without affecting the evaporation process. Here, spectroscopic methods with spatial resolution can discriminate between the components of a liquid mixture. We show that confocal Raman microscopy and spatially resolved NMR spectroscopy can be used as complementary methods to measure concentration gradients in evaporating 1-butanol/1-hexanol droplets on a hydrophobic surface. Deuterating one of the liquids allows analysis of the local composition through the comparison of the intensities of the C­H and C­D stretching bands in Raman spectra. Thus, a concentration gradient in the evaporating droplet was established. Spatially resolved NMR spectroscopy revealed the composition at different positions of a droplet evaporating in the NMR tube, an environment in which air exchange is less pronounced. While not being perfectly comparable, both methods­confocal Raman and spatially resolved NMR experiments­show the presence of a vertical concentration gradient as 1-butanol/1-hexanol droplets evaporate.

Investigation and comparison of resin materials in transparent DLP-printing for application in cell culture and organs-on-a-chip.

Organs-on-a-Chip (OOCs) have recently led to major discoveries and a better understanding of 3D cell organization, cell-cell interactions and tissue response to drugs and biological cues. However, their complexity and variability are still limited by the available fabrication technology. Transparent, cytocompatible and high-resolution 3D-printing could overcome these limitations, offering a flexible and low-cost alternative to soft lithography. Many advances have been made in stereolithography printing regarding resin formulation and the general printing process, but a systematic analysis of the printing process steps, employed resins and post-treatment procedures with a strong focus on the requirements in OOCs is missing. To fill this gap, this work provides an in-depth analysis of three different resin systems in comparison to polystyrene (PS) and poly(dimethylsiloxane) (PDMS), which can be considered the gold-standards in cell culture and microfluidics. The resins were characterized with respect to transparency, cytocompatibility and print resolution. These properties are not only governed by the resin composition, but additionally by the post-treatment procedure. The investigation of the mechanical (elastic modulus ∼2.2 GPa) and wetting properties (∼60° native / 20° plasma treated) showed a behavior very similar to PS. In addition, the absorbance of small molecules was two orders of magnitude lower in the applied resins (diffusion constant ∼0.01 µm2 s-1) than for PDMS (2.5 µm2 s-1), demonstrating the intrinsic suitability of these materials for OOCs. Raman spectroscopy and UV/VIS spectrophotometry revealed that post-treatment increased monomer conversion up to 2 times and removed photo initiator residues, leading to an increased transparency of up to 50% and up to 10-times higher cell viability. High magnification fluorescence imaging of HUVECs and L929 cells cultivated on printed dishes shows the high optical qualities of prints fabricated by the Digital Light Processing (DLP) printer. Finally, components of microfluidic chips such as high-aspect ratio pillars and holes with a diameter of 50 µm were printed. Concluding, the suitability of DLP-printing for OOCs was demonstrated by filling a printed chip with a cell-hydrogel mixture using a microvalve bioprinter, followed by the successful cultivation under perfusion. Our results highlight that DLP-printing has matured into a robust fabrication technology ready for application in extensive and versatile OOC research.

EEG-Microstates Reflect Auditory Distraction After Attentive Audiovisual Perception Recruitment of Cognitive Control Networks.

Processing of sensory information is embedded into ongoing neural processes which contribute to brain states. Electroencephalographic microstates are semi-stable short-lived power distributions which have been associated with subsystem activity such as auditory, visual and attention networks. Here we explore changes in electrical brain states in response to an audiovisual perception and memorization task under conditions of auditory distraction. We discovered changes in brain microstates reflecting a weakening of states representing activity of the auditory system and strengthening of salience networks, supporting the idea that salience networks are active after audiovisual encoding and during memorization to protect memories and concentrate on upcoming behavioural response.

Dataset of oxygen, carbon, and strontium isotope values from the Imperial Roman site of Velia (ca. 1st-2nd c. CE), Italy.

The oxygen (δ18Ocarbonate), strontium (87Sr/86Sr), and previously unpublished carbon (δ13Ccarbonate) isotope data presented herein from the Imperial Roman site of Velia (ca. 1st to 2nd c. CE) were obtained from the dental enamel of human permanent second molars (M2). In total, the permanent M2s of 20 individuals (10 male and 10 female) were sampled at the Museo delle Civiltà in Rome (formerly the Museo Nazionale Preistorico Etnografico "L. Pigorini") and were subsequently processed and analysed at McMaster University. A subsample of teeth (n=5) was initially subjected to Fourier transform infrared spectroscopy (FTIR) analysis to assess for diagenetic alteration through calculation of crystallinity index (CI) values. Subsequently, tooth enamel was analysed for δ13Ccarbonate and δ18Ocarbonate (VPDB) using a VG OPTIMA Isocarb isotope ratio mass spectrometer (IRMS) at McMaster Research for Stable Isotopologues (MRSI), and 87Sr/86Sr was measured by dynamic multi-collection using a thermal ionization mass spectrometer (TIMS) in the School of Geography and Earth Sciences. The dental enamel isotope data presented represent the first δ18O, δ13Ccarbonate, and 87Sr/86Sr values analysed from Imperial Roman Campania to date, providing data of use for comparative analyses of δ18O, δ13C, and 87Sr/86Sr values within the region and for assisting in documenting human mobility in archaeological contexts. Full interpretation of the δ18O and 87Sr/86Sr data presented here is provided in "Imperial Roman mobility and migration at Velia (1st to 2nd c. CE) in southern Italy" [1].

Systematic study of precursor effects on structure and oxygen reduction reaction activity of FeNC catalysts.

In this work, the effect of porphyrin loading and template size is varied systematically to study its impact on the oxygen reduction reaction (ORR) activity and selectivity as followed by rotating ring disc electrode experiments in both acidic and alkaline electrolytes. The structural composition and morphology are investigated by 57Fe Mössbauer spectroscopy, transmission electron microscopy, Raman spectroscopy and Brunauer-Emmett-Teller analysis. It is shown that with decreasing template size, specifically the ORR performance towards fuel cell application gets improved, while at constant area loading of the iron precursor (here expressed in number of porphyrin layers), the iron signature does not change much. Moreover, it is well illustrated that too large area loadings result in the formation of undesired side phases that also cause a decrease in the performance, specifically in acidic electrolyte. Thus, if the impact of morphology is the focus of research it is important to consider the area loading rather than its weight loading. At constant weight loading, beside morphology the structural composition can also change and impact the catalytic performance. This article is part of the theme issue 'Bio-derived and bioinspired sustainable advanced materials for emerging technologies (part 2)'.

Immobilisation of CXCL8 gradients in microfluidic devices for migration experiments.

The release of inflammatory chemokines leads to the formation of chemokine gradients that result in the directed migration of immune cells to the site of injury. In this process, cells respond to soluble gradients (chemotaxis) as well as to immobilised gradients (haptotaxis). Surface-bound chemokine gradients are mostly presented by endothelial cells and supported by glycosaminoglycans (GAGs), such as heparan sulfate, involving the GAG binding site of chemokines. Microfluidic devices have been used to analyse cell migration along soluble chemokine gradients, as these devices allow the generation of stable gradients with resolutions in the range of microns. To immobilise well-controlled soluble gradients of interleukin-8 (CXCL8), an inflammatory chemokine, we developed a simple procedure using a heparin-coated PDMS-microfluidic device. We used these immobilised gradients for migration experiments with CXCL8-responsive THP-1 cells and confirmed directed cell migration. This setup might be useful for the examination of factors that may alter chemotaxis and haptotaxis as well as synergistic and antagonistic effects of other soluble and immobilised chemokines.

Endothelialization of PDMS-based microfluidic devices under high shear stress conditions.

Microfluidic systems made out of polydimethylsiloxane (PDMS) offer a platform to mimic vascular flow conditions in model systems at well-defined shear stresses. However, extracellular matrix (ECM) proteins that are physisorbed on the PDMS are not reliably attached under high shear stress conditions, which makes long-term experiments difficult. To overcome this limitation, we functionalized PDMS surfaces with 3-aminopropyltriethoxysilane (APTES) by using different surface activation methods to develop a stable linkage between the PDMS surface and collagen, which served as a model ECM protein. The stability of the protein coating inside the microfluidic devices was evaluated in perfusion experiments with phosphate-buffered saline (PBS) at 10-40 dynes/cm2 wall shear stress. To assess the stability of cell adhesion, endothelial cells were grown in a multi-shear device over a shear stress range of 20-150 dynes/cm2. Cells on the APTES-mediated collagen coating were stable over the entire shear stress range in PBS (pH 9) for 48 h. The results suggest that at high pH values, the electrostatic interaction between APTES-coated surfaces and collagen molecules offer a very promising tool to modify PDMS-based microfluidic devices for long-term endothelialization under high shear stress conditions.

Probing Charge Accumulation at SrMnO3/SrTiO3 Heterointerfaces via Advanced Electron Microscopy and Spectroscopy.

The last three decades have seen a growing trend toward studying the interfacial phenomena in complex oxide heterostructures. Of particular concern is the charge distribution at interfaces, which is a crucial factor in controlling the interface transport behavior. However, the study of the charge distribution is very challenging due to its small length scale and the intricate structure and chemistry at interfaces. Furthermore, the underlying origin of the interfacial charge distribution has been rarely studied in-depth and is still poorly understood. Here, by a combination of aberration-corrected scanning transmission electron microscopy (STEM) and spectroscopy techniques, we identify the charge accumulation in the SrMnO3 (SMO) side of SrMnO3/SrTiO3 heterointerfaces and find that the charge density attains the maximum of 0.13 ± 0.07 e-/unit cell (uc) at the first SMO monolayer. Based on quantitative atomic-scale STEM analyses and first-principle calculations, we explore the origin of interfacial charge accumulation in terms of epitaxial strain-favored oxygen vacancies, cationic interdiffusion, interfacial charge transfer, and space-charge effects. This study, therefore, provides a comprehensive description of the charge distribution and related mechanisms at the SMO/STO heterointerfaces, which is beneficial for the functionality manipulation via charge engineering at interfaces.

Direct Observation of Huge Flexoelectric Polarization around Crack Tips.

Flexoelectricity is especially relevant for nanoscale structures, and it is expected to be largest at the tip of cracks. We demonstrate the presence of a huge flexoelectric polarization at crack tips in SrTiO3 by direct observation with scanning transmission electron microscopy. We observe an averaged polarization of 62 ± 16 µC cm-2 in the three unit cells adjacent to the crack tip, which is one of the largest flexoelectric polarizations ever reported. The polarization is screened by an electron density of 0.7 ± 0.1 e-/uc localized within one unit cell. These findings reveal the relevance of flexoelectricity for the science of crack formation and propagation.

Insights into the interplay of wetting and transport in mesoporous silica films.

The understanding and design of wetting-transport and wetting-charge-transport interplay in nanometer-sized pores is a still not fully understood key step in improving nanopore transport-related applications. A control of mesopore wettability accompanied by pore filling and ionic mesopore accessibility analysis is expected to deliver major insights into this interplay of nanoscale pore wetting and transport. For a systematic understanding, we demonstrate a gradual adjustment of nanopore ionic accessibility by gradually tuning silica nanopore wettability using chemical vapor phase deposition of 1H,1H,2H,2H-perfluorooctyl dimethylchlorosilane. The mutual influence of wetting on liquid imbibition, condensation, and molecular transport as well as on heat transfer were studied by ellipsometry, cyclic voltammetry and boiling experiments, respectively. A multi-methodical analytic approach was used to directly couple wetting properties of mesoporous silica thin films to ionic mesopore accessibility allowing us to determine two different ion transport mechanisms based on three defined wetting regimes as well as a threshold hydrophobicity suppressing pore accessibility. Furthermore, boiling experiments showed a clear increase in nucleation site density upon changing the wettability of the mesoporous surfaces from hydrophilic to hydrophobic. Hence, these results provide insights into the complex interplay of pore wall functionalization, wetting, and charge-dependent nanopore properties.

The Phenotypic Variation of a Parkin-Related Parkinson's Disease Family and the Role of Heterozygosity.

BACKGROUND: Parkinson's disease (PD) is a common neurodegenerative disorder with both sporadic occurrence and Mendelian heredity, as it is true for autosomal recessive parkin-related PD (PARK-parkin). Parkin-related PD is characterized by early onset, slow progression, frequent lower limb dystonia, and a robust response to levodopa. Clinicians are increasingly confronted with heterozygous PD patients mimicking dominant inheritance. Nevertheless, the exact clinical implications of heterozygosity are not fully understood. CASES: We present an illustrative PARK-parkin family with 2 affected sisters (compound heterozygous) and their father (heterozygous). One sister expresses the classical phenotype, whereas the other has isolated jerky tremor. The father has left-sided action tremor of the hand with some dystonic posturing without clear bradykinesia and normal DaTSCAN. CONCLUSION: This case series illustrates the phenotypic variability in parkin-related PD with 1 classical phenotype and 1 patient with isolated jerky tremor. Unilateral hand tremor of the heterozygous father could mislead genetic testing by mimicking dominant inheritance.

Structural Properties and Magnetic Ordering in 2D Polymer Nanocomposites: Existence of Long Magnetic Dipolar Chains in Zero Field.

The existence of magnetic dipolar nanoparticle chains at zero field has been predicted theoretically for decades, but these structures are rarely observed experimentally. A prerequisite is a permanent magnetic moment on the particles forming the chain. Here we report on the observation of magnetic dipolar chains of spherical iron oxide nanoparticles with a diameter of 12.8 nm. The nanoparticles are embedded in an ultrathin polymer film. Due to the high viscosity of the polymer matrix, the dominating aggregation mechanism is driven by dipolar interactions. Smaller iron oxide nanoparticles (8 nm) show no permanent magnetic moment and do not form chains but compact aggregates. Mixed monolayers of iron oxide nanoparticles and polymer at the air-water interface are characterized by Langmuir isotherms and in situ X-ray reflectometry (XRR). The combination of the particles with a polymer leads to a stable polymer nanocomposite film at the air-water interface. XRR experiments show that nanoparticles are immersed in a thin polymer matrix of 2 nm. Using atomic force microscopy (AFM) on Langmuir-Blodgett films, we measure the lateral distribution of particles in the film. An analysis of single structures within transferred films results in fractal dimensions that are in excellent agreement with 2D simulations.

Oriented crystallization of PEG induced by confinement in cylindrical nanopores: structural and thermal properties.

Nanoporous ion track-etched polycarbonate is ideally suited for the study of confined polymers via small angle X-ray scattering (SAXS) due to the strictly parallel orientation of the pores as well as their uncorrelated lateral distribution. Nanopores with radii ranging from 17 to 213 nm are prepared and coated with SiO2via atomic layer deposition in order to obtain a well-defined and homogeneous surface. A low molecular weight polyethylene glycol (PEG) homopolymer with a semicrystalline lamellar bulk structure is introduced into the nanopores via melt infiltration. At high temperatures SAXS measurements confirm a uniform filling of the pores with amorphous polymer. Upon cooling below the melting point of PEG, a concentrical structure of semicrystalline lamellae is revealed for large pore radii. We introduce models which successfully describe the combined scattering from nanopores and semicrystalline or amorphous PEG inside. DSC measurements of the confined polymer show a decrease of melting temperature and heat of fusion per gram polymer upon reduction of the pore radius and hint at a change in the lamellar configuration.

Vertebral infection in a male individual buried in the monastic cemetery (Cemetery 2) at Ghazali (ca. 670-1270 CE), northern Sudan.

PURPOSE/RESEARCH QUESTION: This article examines pronounced osteoblastic-osteolytic vertebral lesions in a middle adult male (Ghz-2-033), from the Christian Nubian monastic settlement of Ghazali (ca. 670-1270 CE), Sudan, to explore their potential etiology. METHODS: Morphological assessments of sex and age were undertaken in conjunction with macroscopic and radiological methods of assessment for the skeletal lesions documented. RESULTS: Macroscopic assessment of Ghz-2-033 identified mixed osteoblastic-osteolytic lesions in L2-L3 with minor foci in T12-L1, while radiological assessment identified no further lesions. This paleopathological analysis considers tuberculosis, brucellosis, pyogenic intervertebral disc infection, neoplastic conditions, and mycotic infections as potential etiologies. CONCLUSIONS: Tuberculosis is the most probable etiology for the lesions observed. This assessment is based on the morphology of the lesions in conjunction with the known confined living quarters at Ghazali and the presence of tuberculosis vectors (i.e. cattle) in the region. CONTRIBUTIONS TO KNOWLEDGE/ORIGINALITY/VALUE: This brief communication contributes original data documenting the presence of tubercular lesions in a monk buried at the Christian Nubian monastery of Ghazali. On a broader level this study contributes to regional and temporal paleopathological dialogues regarding interactions with pathogens in Christian Nubian monastic contexts. LIMITATIONS FOR THIS STUDY: The potentiality of co-infection with other pathogens (e.g. brucellosis, Staphylococcus) with similar macromorphological traits in skeletal remains cannot be entirely discounted. SUGGESTIONS FOR FURTHER RESEARCH: The use of biomolecular analyses may help to clarify the potential presence of tuberculosis in individual Ghz-2-033.